Gesine Kögler

Publications of Gesine Kögler

• Survival, integration and differentiation of unrestricted somatic stem cells (USSCs) in the heart.

  Authors: Zhaoping Ding, Sandra Burghoff, Anja Buchheiser, Gesine Kögler, Jürgen Schrader

  Cell transplantation. 04/2012;

  A typical human heart infarction involves the loss of approximately 1 billion cardiomyocytes, therefore, many investigators have sought to identify endogenous or exogenous stem cells with theA typical human heart infarction involves the loss of approximately 1 billion cardiomyocytes, therefore, many investigators have sought to identify endogenous or exogenous stem cells with the capacity to differentiate into committed cardiomyocytes and to repopulate lost myocardium. As a result of these efforts, dozens of stem cell types have been reported to have cardiac potential, including pluripotent embryonic stem cells and various adult stem cells from bone marrow, peripheral tissues, and the heart itself. However, despite encouraging results from more than 140 worldwide clinical trials, considerable disagreement exists regarding the longterm efficiency and even the reality of cardiac differentiation by many of these stem cell types, making these issues a continuing source of controversy in the field of stem cell therapy. For donor cell types already in clinical studies, the predominant in vivo effect of bone marrow or endothelial progenitor cells may be on neoangiogenesis and not cardiac specification. Moreover skeletal myoblasts, despite integration and survival, are confounded by arrhythmia, perhaps reflecting lack of transdifferentiation. These obstacles underscore the need to seek cardiac progenitor cells beyond the few known sources, but so far the optimal choice of donor cell is still under intensive research.

• Oxygen tension modifies the 'stemness' of human cord blood-derived stem cells.

  Authors: Anja Buchheiser, Amelie Pia Houben, Julia Bosch, Jendrik Marbach, Stefanie Liedtke, Gesine Kögler

  Cytotherapy. 04/2012;

  Background aims. Amongst different stem cell populations derived from human cord blood (CB), unrestricted somatic stem cells (USSC) are distinguished from CB mesenchymal stromal cells (CB MSC) byBackground aims. Amongst different stem cell populations derived from human cord blood (CB), unrestricted somatic stem cells (USSC) are distinguished from CB mesenchymal stromal cells (CB MSC) by expression patterns of homeobox (HOX) genes, delta-like1 homolog (DLK1) expression and adipogenic differentiation potential. In this study we investigated the effects of oxygen tension on the generation, proliferation and expression of stem cell marker genes, which could be critical during large-scale cell culture for clinical applications. Methods. We cultured CB-derived stem cells at 5% and 20% O(2). Telomere length shortening was analyzed and we investigated gene expression using reverse-transcription (RT)-polymerase chain reaction (PCR) and real-time PCR. Additionally we performed adipogenic and osteogenic in vitro differentiation. Results. Altering the cultivation conditions of USSC or CB MSC from 20% to 5% O(2) had no significant impact. In contrast, cell populations derived from primary cultures prepared at 5% O(2) qualified as neither USSC nor as CB MSC. When converted to 20%, their proliferation was diminished, telomere shortening was accelerated, and two of six cell lines ceased expression of HOX genes. The HOX code of the other cell populations was not been affected by culture conditions. Conclusions. Altering culture conditions during generation can impact cell characteristics such as the HOX code. These effects need to be considered when dealing with cell cultures for clinical applications.

• Impact of DAG stimulation on mineral synthesis, mineral structure and osteogenic differentiation of human cord blood stem cells.

  Authors: Lydia Lammers, Christian Naujoks, Karin Berr, Rita Depprich, Norbert Kübler, Ulrich Meyer, Fabian Langenbach, Beate Lüttenberg, Gesine Kögler, Hans-Peter Wiesmann, Jörg Handschel

  Stem cell research. 03/2012; 8(2):193-205.

  It remains unexplored in what way osteogenic stimulation with dexamethasone, ascorbic acid and β-glycerol phosphate (DAG) influences the process of mineralization, the composition and structure ofIt remains unexplored in what way osteogenic stimulation with dexamethasone, ascorbic acid and β-glycerol phosphate (DAG) influences the process of mineralization, the composition and structure of the assembled mineral. Therefore, we analyzed and characterized biomineralization in DAG-stimulated and unstimulated 3D human unrestricted somatic stem cell (USSC) cultures. Microspheres were analyzed by histological staining, scanning electron microscopy (SEM), semi-quantitative energy-dispersive X-ray spectroscopy (EDX), quantitative wavelength-dispersive X-ray spectroscopy (WDX), transmission electron microscopy (TEM), selected area electron diffraction (SAED) and Raman spectroscopy. Mineral material was detected by SEM and histological staining in both groups, and showed structural differences. DAG influenced the differentiation of USSCs and the formation, structure and composition of the assembled mineral. SEM showed that cells of the +DAG spheres exhibited morphological signs of osteoblast-like cells. EDX and WDX confirmed a Ca-P mineral in both groups. Overall, the mineral material found showed structural similarities to the mineral substance of bony material.

• Improvement of the cell-loading efficiency of biomaterials by inoculation with stem cell-based microspheres, in osteogenesis.

  Authors: Fabian Langenbach, Christian Naujoks, Andrea Laser, Michael Kelz, Pia Kersten-Thiele, Karin Berr, Rita Depprich, Norbert Kübler, Gesine Kögler, Jörg Handschel

  Journal of biomaterials applications. 01/2012; 26(5):549-64.

  In critical-size bone defects, autologous or allogenic cells are required in addition to compatible biomaterials for the successful defect healing. State of the art inoculation methods ofIn critical-size bone defects, autologous or allogenic cells are required in addition to compatible biomaterials for the successful defect healing. State of the art inoculation methods of biomaterials are based on the application of cell suspensions to the biomaterial. However, only less amounts of cells can be applied and sufficient adhesion to the material is required. Therefore, it was investigated whether the advantages of stem cell-based microspheres and insoluble collagenous bone matrix (ICBM) scaffolds can be combined which can lead to an advancement in cell seeding on biomaterials. Microspheres were produced from unrestricted somatic stem cells from human umbilical cord blood and were mounted on ICBM scaffolds. Following the incubation with osteogenic or control medium, the constructs were analyzed histologically after 3, 7, 14, and 28 days. Alizarin Red S and von Kossa staining revealed microsphere mineralization after 3 days in osteogenic and after 14 days in control medium. Meanwhile, a time-dependent increase in tissue, growing out of the microspheres, was detected. Our results provide evidence that microsphere-ICBM constructs are promising candidates for approaches of bone regeneration. They allow the transfer of substantially high numbers of cells in partially mineralized constructs.

• Distinct Differentiation Potential of "MSC" Derived from Cord Blood and Umbilical Cord: Are Cord-Derived Cells True Mesenchymal Stromal Cells?

  Authors: Julia Bosch, Amelie Pia Houben, Teja Falk Radke, Daniela Stapelkamp, Erich Bünemann, Percy Balan, Anja Buchheiser, Stefanie Liedtke, Gesine Kögler

  Stem cells and development. 11/2011;

  Mesenchymal stromal cells (MSC) with distinct differentiation properties have been reported in many adult [eg, bone marrow (BM)] or fetal tissues [eg, cord blood (CB); umbilical cord (UC)] and areMesenchymal stromal cells (MSC) with distinct differentiation properties have been reported in many adult [eg, bone marrow (BM)] or fetal tissues [eg, cord blood (CB); umbilical cord (UC)] and are defined by their specific surface antigen expression and multipotent differentiation potential. The MSC identity of these cells should be validated by applying well-defined readout systems if a clinical application is considered. In order to determine whether cells isolated from human UC fulfill the criteria defined for MSC, the immunophenotype and differentiation potential including gene expression analysis of the most relevant lineage-specific markers were analyzed in the presented report in combination with the HOX-gene expression. Cells from the UC do not differentiate into osteoblasts demonstrated by Alizarin Red and Von Kossa staining in addition to real-time polymerase chain reaction (PCR)-analysis of runt-related transcription factor 2, bone sialoprotein, osteocalcin, osterix, bone morphogenetic proteins 2 and 4. Oil Red O staining as well as PCR analysis of peroxisome proliferator-activated receptor-gamma, fatty acid-binding protein 4, and perilipin revealed an absent adipogenic differentiation. The lack of potential to differentiate into chondrocytes was documented by Alcian-Blue periodic acid-Schiff, Safranin O staining, and real-time PCR analysis of SOX9. Furthermore, neither endothelial nor myogenic differentiation was documented after induction of UC-MSC. In comparison to CB- and BM-derived cells, UC cells revealed an absent trilineage differentiation capacity in vitro. Therefore, these cells should not be termed "mesenchymal stromal cells." The UC cells can be distinguished from CB- and BM-derived cells as well as from pericytes and foreskin fibroblasts by the expression of HOX-genes and the cell surface antigens CD56 and CD146.

• Generation and differentiation of microtissues from multipotent precursor cells for use in tissue engineering.

  Authors: Fabian Langenbach, Karin Berr, Christian Naujoks, Andrea Hassel, Michael Hentschel, Rita Depprich, Norbert R Kubler, Ulrich Meyer, Hans-Peter Wiesmann, Gesine Kögler, Jörg Handschel

  Nature protocols. 11/2011; 6(11):1726-35.

  This protocol describes an effective method for the production of spherical microtissues (microspheres), which can be used for a variety of tissue-engineering purposes. The obtained microtissues areThis protocol describes an effective method for the production of spherical microtissues (microspheres), which can be used for a variety of tissue-engineering purposes. The obtained microtissues are well suited for the study of osteogenesis in vitro when multipotent stem cells are used. The dimensions of the microspheres can easily be adjusted according to the cell numbers applied in an individual experiment. Thus, microspheres allow for the precise administration of defined cell numbers at well-defined sites. Here we describe a detailed workflow for the production of microspheres using unrestricted somatic stem cells from human umbilical cord blood and adapted protocols for the use of these microspheres in histological analysis. RNA extraction methods for mineralized microtissues are specifically modified for optimum yields. The duration of running the complete protocol without preparatory cell culture but including 2 weeks of microsphere incubation, histological staining and RNA isolation is about 3 weeks.

• Significant clinical, neuropathological and behavioural recovery from acute spinal cord trauma by transplantation of a well-defined somatic stem cell from human umbilical cord blood.

  Authors: Jessica Schira, Marcia Gasis, Veronica Estrada, Marion Hendricks, Christine Schmitz, Thorsten Trapp, Fabian Kruse, Gesine Kögler, Peter Wernet, Hans-Peter Hartung, Hans Werner Müller

  Brain : a journal of neurology. 09/2011; 135(Pt 2):431-46.

  Stem cell therapy is a potential treatment for spinal cord injury and different stem cell types have been grafted into animal models and humans suffering from spinal trauma. Due to inconsistentStem cell therapy is a potential treatment for spinal cord injury and different stem cell types have been grafted into animal models and humans suffering from spinal trauma. Due to inconsistent results, it is still an important and clinically relevant question which stem cell type will prove to be therapeutically effective. Thus far, stem cells of human sources grafted into spinal cord mostly included barely defined heterogeneous mesenchymal stem cell populations derived from bone marrow or umbilical cord blood. Here, we have transplanted a well-defined unrestricted somatic stem cell isolated from human umbilical cord blood into an acute traumatic spinal cord injury of adult immune suppressed rat. Grafting of unrestricted somatic stem cells into the vicinity of a dorsal hemisection injury at thoracic level eight resulted in hepatocyte growth factor-directed migration and accumulation within the lesion area, reduction in lesion size and augmented tissue sparing, enhanced axon regrowth and significant functional locomotor improvement as revealed by three behavioural tasks (open field Basso-Beattie-Bresnahan locomotor score, horizontal ladder walking test and CatWalk gait analysis). To accomplish the beneficial effects, neither neural differentiation nor long-lasting persistence of the grafted human stem cells appears to be required. The secretion of neurite outgrowth-promoting factors in vitro further suggests a paracrine function of unrestricted somatic stem cells in spinal cord injury. Given the highly supportive functional characteristics in spinal cord injury, production in virtually unlimited quantities at GMP grade and lack of ethical concerns, unrestricted somatic stem cells appear to be a highly suitable human stem cell source for clinical application in central nervous system injuries.

• Neonatal NK-cell repertoires are functionally, but not structurally, biased toward recognition of self HLA class I.

  Authors: Kathrin Schönberg, Johannes C Fischer, Gesine Kögler, Markus Uhrberg

  Blood. 03/2011; 117(19):5152-6.

  Human natural killer (NK)-cell repertoires are biased toward more frequent expression of inhibitory killer cell Ig-like receptor (KIR) receptors for self-human leukocyte antigen (HLA) class I.Human natural killer (NK)-cell repertoires are biased toward more frequent expression of inhibitory killer cell Ig-like receptor (KIR) receptors for self-human leukocyte antigen (HLA) class I. Moreover, only those NK cells that express cognate receptors for self are fully functional in terms of cytotoxicity and cytokine production. It is so far unknown whether functional education and structural adaptation to HLA class I are implemented during NK-cell development and whether both processes are mechanistically connected. Here we show that NK-cell repertoires in cord blood are not yet shaped toward increased clonal frequencies of KIR for self-HLA class I as determined for the 3 major KIR ligands C1, C2, and Bw4. Nonetheless, neonatal NK cells expressing cognate KIR exhibited enhanced effector function on the level of degranulation and cytokine production. The study suggests that functional education of cognate KIR by self-HLA class I precedes structural adaptation of KIR repertoires and that both processes are not directly linked to each other.

• Unrestricted somatic stem cells: interaction with CD34+ cells in vitro and in vivo, expression of homing genes and exclusion of tumorigenic potential.

  Authors: Kathrin Sonja Jeltsch, Teja Falk Radke, Stephanie Laufs, Frank Anton Giordano, Heike Allgayer, Frederik Wenz, Walter Jens Zeller, Gesine Kögler, Stefan Fruehauf, Patrick Maier

  Cytotherapy. 10/2010; 13(3):357-65.

  Transplantation of allogeneic hematopoietic stem cells (HSC) within the framework of hematologic oncology or inherited diseases may be associated with complications such as engraftment failure andTransplantation of allogeneic hematopoietic stem cells (HSC) within the framework of hematologic oncology or inherited diseases may be associated with complications such as engraftment failure and long-term pancytopenia. HSC engraftment can be improved, for example by co-transplantation with mesenchymal stem cells (MSC). Recently, a new multipotent MSC line from umbilical cord blood, unrestricted somatic stem cells (USSC), has been described. It was demonstrated that USSC significantly support proliferation of HSC in an in vitro feeder layer assay. A NOD/SCID mouse model was used to assess the effect of USSC on co-transplanted CD34(+) cells and look for the fate of transplanted USSC. The migration potential of USSC was studied in a Boyden chamber migration assay and in vivo. Quantitative real-time polymerase chain reaction (qRT-PCR) for CXCR4, CD44, LFA1, CD62L, VLA4, RAC2, VLA5A and RAC1 were performed. NMR1 nu/nu mice were used for a tumorigenicity test. After 4 weeks, homing of human cells (CD45(+)) to the bone marrow of NOD/SCID mice was significantly increased in mice co-transplanted with CD34(+) cells and USSC (median 30.9%, range 7-50%) compared with the CD34(+) cell-only control group (median 5.9%, range 3-10%; P = 0.004). Homing of USSC could not be shown in the bone marrow. A cell-cell contact was not required for the graft enhancing effect of USSC. An in vivo tumorigenicity assay showed no tumorigenic potential of USSC. This pre-clinical study clearly shows that USSC have an enhancing effect on engraftment of human CD34(+) cells. USSC are a safe graft adjunct.

• Induction of pluripotency in human cord blood unrestricted somatic stem cells.

  Authors: Holm Zaehres, Gesine Kögler, Marcos J Arauzo-Bravo, Martina Bleidissel, Simeon Santourlidis, Sandra Weinhold, Boris Greber, Jeong Beom Kim, Anja Buchheiser, Stefanie Liedtke [......] Xiaoyi Zhao, Johann Meyer, Peter Reinhardt, Boris Burr, Simon Waclawczyk, Claudia Ortmeier, Markus Uhrberg, Hans R Schöler, Tobias Cantz, Peter Wernet

  Experimental hematology. 09/2010; 38(9):809-18, 818.e1-2.

  Generation of induced pluripotent stem (iPS) cells from human cord blood (CB)-derived unrestricted somatic stem cells and evaluation of their molecular signature and differentiation potential inGeneration of induced pluripotent stem (iPS) cells from human cord blood (CB)-derived unrestricted somatic stem cells and evaluation of their molecular signature and differentiation potential in comparison to human embryonic stem cells. Unrestricted somatic stem cells isolated from human CB were reprogrammed to iPS cells using retroviral expression of the transcription factors OCT4, SOX2, KLF4, and C-MYC. The reprogrammed cells were analyzed morphologically, by quantitative reverse transcription polymerase chain reaction, genome-wide microRNA and methylation profiling, and gene expression microarrays, as well as in their pluripotency potential by in vivo teratoma formation in severe combined immunodeficient mice and in vitro differentiation. CB iPS cells are very similar to human embryonic stem cells morphologically, at their molecular signature, and in their differentiation potential. Human CB-derived unrestricted somatic stem cells offer an attractive source of cells for generation of iPS cells. Our findings open novel perspectives to generate human leukocyte antigen-matched pluripotent stem cell banks based on existing CB banks. Besides the obvious relevance of a second-generation CB iPS cell bank for pharmacological and toxicological testing, its application for autologous or allogenic regenerative cell transplantation appears feasible.

• The HOX Code as a "biological fingerprint" to distinguish functionally distinct stem cell populations derived from cord blood.

  Authors: Stefanie Liedtke, Anja Buchheiser, Julia Bosch, Frank Bosse, Fabian Kruse, Xiaoyi Zhao, Simeon Santourlidis, Gesine Kögler

  Stem cell research. 07/2010; 5(1):40-50.

  Mesenchymal stem cells (MSC) have been isolated from almost every adult tissue. In cord blood (CB), different non-hematopoietic CD45-, CD34- adherent cell populations can be generated: the cord bloodMesenchymal stem cells (MSC) have been isolated from almost every adult tissue. In cord blood (CB), different non-hematopoietic CD45-, CD34- adherent cell populations can be generated: the cord blood derived MSC (CB-MSC), that behave almost like MSC from bone marrow (BM-MSC), and unrestricted somatic stem cells (USSC) which show a distinct differentiation potential into all three germ layers. However, distinguishing these populations easily by molecular markers is still a concern. In this study we were able to present the HOX expression pattern of USSC, CB-MSC and BM-MSC, which in fact allows a discrimination of these populations. Briefly, RT-PCR analysis of the HOX code revealed a high similarity between BM-MSC and CB-MSC, which are both HOX-positive, whereas USSC resembled H9 embryonic stem cells HOX-negative.Especially HOXA9, HOXB7, HOXC10 and HOXD8 are good candidate markers to discriminate MSC from USSC. Thus, our data suggest that the "biological fingerprint" based on the HOX code can be used to distinguish functionally distinct MSC populations derived from bone marrow and cord blood.

• Biocompatibility of osteogenic predifferentiated human cord blood stem cells with biomaterials and the influence of the biomaterial on the process of differentiation.

  Authors: Christian Naujoks, Fabian Langenbach, Karin Berr, Rita Depprich, Norbert Kübler, Ulrich Meyer, Jörg Handschel, Gesine Kögler

  Journal of biomaterials applications. 03/2010; 25(5):497-512.

  Modern cell-based bone reconstruction therapies offer new therapeutic opportunities and tissue engineering represents a more biological-oriented approach to heal bone defects of the skeleton. HumanModern cell-based bone reconstruction therapies offer new therapeutic opportunities and tissue engineering represents a more biological-oriented approach to heal bone defects of the skeleton. Human unrestricted somatic stem cells (USSCs) derived form umbilical cord blood offer new promising aspects e.g., can differentiate into osteogenetic cells. Furthermore these cells have fewer ethical and legal restrictions compared to embryonic stem cells (ESCs). The purpose of this study was to evaluate the compatibility of osteogenic pre-differentiated USSCs with various biomaterials and to address the question, whether biomaterials influence the process of differentiation of the USSCs. After osteogenic differentiation with DAG USSCs were cultivated with various biomaterials. To asses the biocompatibility of USSCs the attachment and the proliferation of the cells on the biomaterial were measured by a CyQUANT(®) assay, the morphology was analyzed by scanning electron microscopy and the influence of the gene expression was analyzed by real time PCR. Our results provide evidence that insoluble collagenous bone matrix followed by β-tricalciumphosphate is highly suitable for bone tissue engineering regarding cell attachment and proliferation. The gene expression analysis indicates that biomaterials influence the gene expression of USSCs. These results are in concordance with our previous study with ESCs.

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• Comparison of ectopic bone formation of embryonic stem cells and cord blood stem cells in vivo.

  Authors: Jörg Handschel, Christian Naujoks, Fabian Langenbach, Karin Berr, Rita A Depprich, Michelle A Ommerborn, Norbert R Kübler, Matthias Brinkmann, Gesine Kögler, Ulrich Meyer

  Tissue engineering. Part A. 03/2010; 16(8):2475-83.

  Cell-based reconstruction therapies promise new therapeutic opportunities for bone regeneration. Unrestricted somatic stem cells (USSC) from cord blood and embryonic stem cells (ESCs) can beCell-based reconstruction therapies promise new therapeutic opportunities for bone regeneration. Unrestricted somatic stem cells (USSC) from cord blood and embryonic stem cells (ESCs) can be differentiated into osteogenic cells. The purpose of this in vivo study was to compare their ability to induce ectopic bone formation in vivo. Human USSCs and murine ESCs were cultured as both monolayer cultures and micromasses and seeded on insoluble collagenous bone matrix (ICBM). One week and 1, 2, and 3 months after implanting the constructs in immune-deficient rats, computed tomography scans were performed to detect any calcification. Subsequently, the implanted constructs were examined histologically. The radiological examination showed a steep increase in the mineralized bone-like tissue in the USSC groups. This increase can be considered as statistically significant compared to the basic value. Moreover, the volume and the calcium portion measured by computed tomography scans were about 10 times higher than in the ESC group. The volume of mineralization in the ESC group increased to a much smaller extent over the course of time, and the control group (ICBM without cells) showed almost no alterations during the study. The histological examinations parallel the radiological findings. Cord blood stem cells in combination with ICBM-induced ectopic bone formation in vivo are stronger than ESCs.

• DLK-1 as a marker to distinguish unrestricted somatic stem cells and mesenchymal stromal cells in cord blood.

  Authors: Simone Maria Kluth, Anja Buchheiser, Amelie Pia Houben, Stefanie Geyh, Thomas Krenz, Teja Falk Radke, Constanze Wiek, Helmut Hanenberg, Petra Reinecke, Peter Wernet, Gesine Kögler

  Stem cells and development. 03/2010; 19(10):1471-83.

  In addition to hematopoietic stem cells, cord blood (CB) also contains different nonhematopoietic CD45-, CD34- adherent cell populations: cord blood mesenchymal stromal cells (CB MSC) that behaveIn addition to hematopoietic stem cells, cord blood (CB) also contains different nonhematopoietic CD45-, CD34- adherent cell populations: cord blood mesenchymal stromal cells (CB MSC) that behave almost like MSC from bone marrow (BM MSC) and unrestricted somatic stem cells (USSC) that differentiate into cells of all 3 germ layers. Distinguishing between these populations is difficult due to overlapping features such as the immunophenotype or the osteogenic and chondrogenic differentiation pathway. Functional differences in the differentiation potential suggest different developmental stages or different cell populations. Here we demonstrate that the expression of genes and the differentiation toward the adipogenic lineage can discriminate between these 2 populations. USSC, including clonal-derived cells lacking adipogenic differentiation, strongly expressed δ-like 1/preadipocyte factor 1 (DLK-1/PREF1) correlating with high proliferative potential, while CB MSC were characterized by a strong differentiation toward adipocytes correlating with a weak or negative DLK-1/PREF1 expression. Constitutive overexpression of DLK-1/PREF1 in CB MSC resulted in a reduced adipogenic differentiation, whereas silencing of DLK-1 in USSC resulted in adipogenic differentiation.

• Stem cells derived from cord blood in transplantation and regenerative medicine.

  Authors: Verena Reimann, Ursula Creutzig, Gesine Kögler

  Deutsches Ärzteblatt international. 12/2009; 106(50):831-6.

  Physicians of any specialty may be the first persons to whom prospective parents turn for information about the acquisition and storage of stem cells derived from cord blood. Stem cells canPhysicians of any specialty may be the first persons to whom prospective parents turn for information about the acquisition and storage of stem cells derived from cord blood. Stem cells can potentially be used to treat many diseases, yet they are not a panacea. This article provides an overview of their current and possible future applications. Original papers were retrieved by a selective search of the literature, and the Internet sites and advertising brochures of private stem cell banks were also examined. Allogeneic hematopoietic stem cells derived from umbilical cord blood (obtained from healthy donors, rather than from the patient to be treated) have been in routine use worldwide for more than ten years in the treatment of hematopoietic diseases. Experiments in cell culture and in animal models suggest that these cells might be of therapeutic use in regenerative medicine, but also show that this potential can be realized only if the cells are not cryopreserved. There is as yet no routine clinical application of autologous hematopoietic stem cells from cord blood (self-donation of blood), even though cord blood has been stored in private banks for more than ten years. Autologous stem cells from cord blood have poor prospects for use in regenerative medicine, because they have to be cryopreserved until use. Physicians should tell prospective parents that they have no reason to feel guilty if they choose not to store cord blood in a private bank.

• Osteogenic differentiation influence stem cell migration out of scaffold free microspheres.

  Authors: Fabian Langenbach, Christian Naujoks, Pia Valeska Kersten-Thiele, Karin Berr, Rita Depprich, Norbert Kübler, Gesine Kögler, Jörg Handschel

  Tissue engineering. Part A. 09/2009;

  Complete bone regeneration of critical size defects (CSDs) frequently fail due to the use of acellular bone substitutes and due to partially negative influences of artificial scaffolds. However, theComplete bone regeneration of critical size defects (CSDs) frequently fail due to the use of acellular bone substitutes and due to partially negative influences of artificial scaffolds. However, the supply of cells to CSDs is essential for the regeneration. Therefore, engineered scaffold free tissues, with outgrowing cells that fill up spaces in the bony defect, are promising candidates for bone regeneration approaches. Here we demonstrate such a scaffold free tissue construct (microspheres) that, if osteogenic differentiated, mineralizes while maintaining the capability to let cells grow out of the united cell structure. A superior outgrowth capability of microspheres composed of human cord-blood derived USSCs (unrestricted somatic stem cells) compared to murine ESCs (embryonic stem cells) was found and a time dependent reduction in outgrowth was evident, in vitro. Even after five days of osteoinduction and strong mineralization cells migrate out of the microsphere. Since migration of cells out of USSC-microspheres was also found in extracellular matrix gel, we suggest that cells would migrate also in vivo. Thus microspheres could serve as the scaffold and the source of osteogenic cells in future bone regeneration approaches. Furthermore microspheres permit the precise administration of large amount of cells into an area of interest.

• Cord blood for tissue regeneration.

  Authors: Anja Buchheiser, Stefanie Liedtke, Leendert H J Looijenga, Gesine Kögler

  Journal of cellular biochemistry. 09/2009;

  Umbilical cord blood (CB) has become a commonly accepted source of hematopoietic stem cells for transplantation in children and adults. It is readily available and outperforms bone marrow (BM) asUmbilical cord blood (CB) has become a commonly accepted source of hematopoietic stem cells for transplantation in children and adults. It is readily available and outperforms bone marrow (BM) as well as peripheral blood stem cells in terms of tolerance for HLA-mismatches between donor and recipient and its decreased graft-versus-host disease. Clinical use has been expanded from hematological malignancies to various areas such as treatment of metabolic genetic disorders or to induce angiogenesis. For the last years CB has been under intense experimental investigation in in vitro differentiation models as well as in preclinical animal models. Since CB-derived stem cells offer multiple advantages over adult stem cells from other sources like BM, CB may provide a future source of stem cells for tissue repair and regeneration. To facilitate the use of CB-derived stem cells in clinical scenarios, the biology of these cells needs to be further explored in detail particularly with regard to the fact that different non-hematopoietic stem cell populations occur within CB. Here we explore the most consistent and the most contradictory data referring to the differentiation potential of CB-derived stem cells and give an outlook on their potential clinical value including and possible reprogramming into IPS cells. J. Cell. Biochem. (c) 2009 Wiley-Liss, Inc.

• Hepatocyte growth factor/c-MET axis-mediated tropism of cord blood-derived unrestricted somatic stem cells for neuronal injury.

  Authors: Thorsten Trapp, Gesine Kögler, Abdelouahid El-Khattouti, Rüdiger V Sorg, Michael Besselmann, Melanie Föcking, Christian P Bührle, Ingo Trompeter, Johannes C Fischer, Peter Wernet

  The Journal of biological chemistry. 11/2008; 283(47):32244-53.

  An under-agarose chemotaxis assay was used to investigate whether unrestricted somatic stem cells (USSC) that were recently characterized in human cord blood are attracted by neuronal injury inAn under-agarose chemotaxis assay was used to investigate whether unrestricted somatic stem cells (USSC) that were recently characterized in human cord blood are attracted by neuronal injury in vitro. USSC migrated toward extracts of post-ischemic brain tissue of mice in which stroke had been induced. Moreover, apoptotic neurons secrete factors that strongly attracted USSC, whereas necrotic and healthy neurons did not. Investigating the expression of growth factors and chemokines in lesioned brain tissue and neurons and of their respective receptors in USSC revealed expression of hepatocyte growth factor (HGF) in post-ischemic brain and in apoptotic but not in necrotic neurons and of the HGF receptor c-MET in USSC. Neuronal lesion-triggered migration was observed in vitro and in vivo only when c-MET was expressed at a high level in USSC. Neutralization of the bioactivity of HGF with an antibody inhibited migration of USSC toward neuronal injury. This, together with the finding that human recombinant HGF attracts USSC, document that HGF signaling is necessary for the tropism of USSC for neuronal injury. Our data demonstrate that USSC have the capacity to migrate toward apoptotic neurons and injured brain. Together with their neural differentiation potential, this suggests a neuroregenerative potential of USSC. Moreover, we provide evidence for a hitherto unrecognized pivotal role of the HGF/c-MET axis in guiding stem cells toward brain injury, which may partly account for the capability of HGF to improve function in the diseased central nervous system.

• Suppression of Cellular Immunity by Cord Blood-Derived Unrestricted Somatic Stem Cells is Cytokine Dependent.

  Authors: Meike Winter, Xiao-Nong Wang, Walter Däubener, Annette Eyking, Michelle Rae, Anne M Dickinson, Peter Wernet, Gesine Kögler, Rüdiger V Sorg

  Journal of cellular and molecular medicine. 11/2008;

  Abstract Unrestricted somatic stem cells (USSC) have the potential to differentiate into tissues derived from all three germinal layers and therefore hold promise for use in regenerative therapies.Abstract Unrestricted somatic stem cells (USSC) have the potential to differentiate into tissues derived from all three germinal layers and therefore hold promise for use in regenerative therapies. Furthermore, they have haematopoietic stromal activity, a characteristic which may be exploited to enhance haematopoietic engraftment. Both applications may require USSC to be used in an allogeneic, HLA-mismatched setting. We have therefore studied their in vitro interaction with cellular immunity. USSC showed no allostimulatory activity and caused only minimal inhibition of allogeneic T-cell responses. However, following pre-stimulation with IFNgamma and TNFalpha, they inhibited T-cell proliferation in an indoleamine 2, 3-dioxygenase-dependent manner and suppressed graft-versus-host type reactions. In addition, USSC inhibited DC maturation and function. This inhibition was overridden by stronger DC maturation signals provided by IL-1beta, IL-6, PGE(2) and TNFalpha compared to TNFalpha alone. Pre-stimulation of USSC with IFNgamma and TNFalpha had a similar effect: Inhibition of DC maturation was no longer observed. Thus, USSC are conditionally immunosuppressive, and IFNgamma and TNFalpha constitute a switch, which regulates their immunological properties. They either suppress T-cell responses in the presence of both cytokines or in their absence block DC differentiation and function. These activities may contribute to fine-tuning the immune system especially at sites of tissue damage in order to ensure appropriate differentiation of USSC and subsequent tissue repair. Therapeutically, they may help to protect USSC and possibly their progeny from immune rejection.

• Oct4 expression revisited: potential pitfalls for data misinterpretation in stem cell research.

  Authors: Stefanie Liedtke, Milaid Stephan, Gesine Kögler

  Biological chemistry. 07/2008; 389(7):845-50.

  The octamer-binding transcription factor 4 gene encodes a nuclear protein (Oct4, also known as Pou5F1 and Oct3/4) that belongs to a family of transcription factors containing the POU DNA-bindingThe octamer-binding transcription factor 4 gene encodes a nuclear protein (Oct4, also known as Pou5F1 and Oct3/4) that belongs to a family of transcription factors containing the POU DNA-binding domain. Expression can be detected in embryonic stem cells as well as in adult stem cells, such as bone marrow-derived mesenchymal stem cells. Expression of Oct4 is downregulated coincident with stem cell differentiation and loss of expression leading to differentiation. A role for maintaining pluripotency and self-renewal of embryonic stem cells is ascribed to Oct4 as a pluripotency marker. Results describing Oct4 expression in differentiated cells, including peripheral blood mononuclear cells (PBMCs), neonatal and adult stem cells, as well as cancer cells, must be interpreted with caution. In several publications, Oct4 has been ascribed a function in maintaining self-renewal of adult stem cells. In contrast, other publications reported Oct4 expression in human tumor cells. Here, we summarize the recent findings on Oct4 expression and present possibilities and reasons why several false positive results on Oct4 expression still occur in the recent literature. Also, simple solutions are provided to avoid these positive signals.