Tissue Engineering Laboratory and Berlin-Brandenburg Center for Regenerative Therapies, Department of Rheumatology and Clinical Immunology, Charité-Universitätsmedizin Berlin, CCM, Tucholskystr. 2, 10117 Berlin, Germany.
Recruitment of mesenchymal stem cells (MSC) to tissue damages is a promising approach for in situ tissue regeneration. The physiological mechanisms and regulatory processes of MSC trafficking to injured tissue remain poorly understood. However, the pivotal role of chemokines in MSC recruitment has already been shown. The aim of this study was to determine the migratory potential and the gene expression profile of MSC stimulated with the CC chemokine CCL25 (TECK). Bone marrow derived human MSC were exposed to different doses of CCL25 in a standardized chemotaxis assay. Microarray gene expression profiling and pathway analysis were performed for CCL25 stimulated MSC. Maximum migration of MSC towards CCL25 was observed at 10(3) nM. Microarray analysis revealed an induction of molecules directly involved in chemotaxis and homing of bone marrow cells (CXCL1-3, CXCL8, PDE4B), cytoskeletal and membrane reorganisation (CXCL8, PLD1, IGFBP1), cellular polarity (PLD1), and cell movement (CXCL1-3, CXCL6, CXCL8, PTGS2, PDE4B, TGM2). Respective chemokine secretion was confirmed by protein membrane-array analysis. The activation of CXCR2 ligands (CXCL1-3, CXCL5-6, CXCL8) and a LIF-receptor/gp130 ligand (LIF) indicated an involvement of the respective signaling pathways during initiation of chemotaxis and migration. These results suggest CCL25 as a new potential candidate for further in situ regeneration approaches.
"The increased expression levels of CXCL1 and CXCL6 detected in ADHLSC are in accordance with the parallel up-regulated level of IL-8 and LIF. All these cytokines are directly involved in chemotaxis, cell movement and migration of mesenchymal stem cells . "
[Show abstract][Hide abstract] ABSTRACT: Adult-derived human liver stem/progenitor cells (ADHLSC) are obtained after primary culture of the liver parenchymal fraction. The cells are of fibroblastic morphology and exhibit a hepato-mesenchymal phenotype. Hepatic stellate cells (HSC) derived from the liver non-parenchymal fraction, present a comparable morphology as ADHLSC. Because both ADHLSC and HSC are described as liver stem/progenitor cells, we strived to extensively compare both cell populations at different levels and to propose tools demonstrating their singularity.
ADHLSC and HSC were isolated from the liver of four different donors, expanded in vitro and followed from passage 5 until passage 11. Cell characterization was performed using immunocytochemistry, western blotting, flow cytometry, and gene microarray analyses. The secretion profile of the cells was evaluated using Elisa and multiplex Luminex assays.
Both cell types expressed α-smooth muscle actin, vimentin, fibronectin, CD73 and CD90 in accordance with their mesenchymal origin. Microarray analysis revealed significant differences in gene expression profiles. HSC present high expression levels of neuronal markers as well as cytokeratins. Such differences were confirmed using immunocytochemistry and western blotting assays. Furthermore, both cell types displayed distinct secretion profiles as ADHLSC highly secreted cytokines of therapeutic and immuno-modulatory importance, like HGF, interferon-γ and IL-10.
Our study demonstrates that ADHLSC and HSC are distinct liver fibroblastic cell populations exhibiting significant different expression and secretion profiles.
PLoS ONE 01/2014; 9(1):e86137. DOI:10.1371/journal.pone.0086137 · 3.23 Impact Factor
"Isolation of human/rat MSCs The human bone marrow aspirates were received in accordance with the ethical committee of the Charit e Universitätsmedizin Berlin. hMSCs were cultivated as described previously in Dulbecco's modified Eagle's medium (DMEM, Biochrom, Berlin, Germany) containing 2 ng/mL basic fibroblast growth factor (bFGF, Peprotech, Hamburg, Germany), 10% foetal bovine serum (FBS, Hyclone, Bonn, Germany), 100 mg/mL streptomycin and 100 U/mL penicillin (Biochrom), 20 mM 4-(2-hydroxyethyl)-1-piperazineetha- nesulphonic acid (HEPES)-buffered saline (Biochrom) and 2 mM L-glutamine (Biochrom) (Ringe et al., 2007; Binger et al., 2009). Medium was exchanged after 72 h and every 2–3 days. "
[Show abstract][Hide abstract] ABSTRACT: Chitosan-beta glycerophosphate-hydroxyethyl cellulose (CH-GP-HEC) is a biocompatible and biodegradable scaffold exhibiting a sol-gel transition at 37°C. Chondrogenic factors or mesenchymal stem cells (MSCs) can be included in the CH-GP-HEC, and injected into the site of injury to fill the cartilage tissue defects with minimal invasion and pain. The possible impact of the injectable CH-GP-HEC on the viability of the encapsulated MSCs was assessed by propidium iodide-fluorescein diacetate (PI-FDA) staining. Proliferation of the human and rat MSCs was also determined by MTS assay on days 0, 7, 14, and 28 after encapsulation. To investigate the potential application of CH-GP-HEC as a drug delivery device, the in vitro release profile of insulin was quantified by QuantiPro-BCA(TM) protein assay. Chondrogenic differentiation capacity of the encapsulated human MSCs (hMSCs) was also determined after induction of differentiation with transforming growth factor β3 (TGF-β3). MSCs have very good survival and proliferative rates within CH-GP-HEC hydrogel during the 28 day investigation. A sustained release of insulin occurred over 8 days. The CH-GP-HEC hydrogel also provided suitable conditions for chondrogenic differentiation of the encapsulated hMSCs. In conclusion the high potential of CH-GP-HEC as an injectable hydrogel for cartilage tissue engineering is emphasised.
Cell Biology International 01/2014; 38(1). DOI:10.1002/cbin.10181 · 1.93 Impact Factor
"Obviously, to understand the underlying mechanism would be of interest. In this context, CCL25 has been suggested as a potential chemoattractant for the directional movement of MSCs , and C-C chemokine receptor type 9 (CCR9) is known as a cognate receptor of CCL25 [57,58]. To check whether the chondrogenic differentiated state of MSCs affects the cell-migration rate, we performed the chemotaxis assay for undifferentiated, chondrogenic differentiated and dedifferentiated cells, by using the chemokine CCL25 . "
[Show abstract][Hide abstract] ABSTRACT: Guided migration of chondrogenically differentiated cells has not been well studied before, even though it may be critical for growth, repair and regenerative processes. The chemokine CCL25 is believed to play a critical role in the directional migration of leukocytes and stem cells. To investigate the motility affect of serum or CCL25 mediated chemotaxis on chondrogenically differentiated cells, mesenchymal stem cells (MSC) were induced to chondrogenic lineage cells.
MSC-derived chondrogenically differentiated cells were characterized for morphology, histology, immunohistochemistry, qPCR, surface profile and serum or CCL25 mediated cell migration. Additionally, the chemokine receptor, CCR9, was examined in different states of MSC.
The chondrogenic differentiated state of MSC was positive for collagen type II and alcian blue staining, and showed significantly upregulated expression of COL2A1and SOX9, and downregulated expression of CD44, CD73, CD90, CD105 and CD166, in contrast to undifferentiated and dedifferentiated state of MSC. For chondrogenic differentiated, undifferentiated and dedifferentiated state of MSC, the serum mediated chemotaxis was in % ratio of 33:84:85, and CCL25 mediated chemotaxis was in % ratio of 12:14:13 respectively. On protein level, CCR9, receptor of CCL25, was expressed in the form of extracellular and intracellular domains. On gene level, qPCR confirmed the expression of CCR9 in different states of MSC.
CCL25 is an effective cue to guide migration in a directional way. In CCL25 mediated chemotaxis, the cell migration rate was almost the same for different states of MSC. In serum mediated chemotaxis, the cell migration rate of chondrogenically differentiated cells was significantly lower than undifferentiated or dedifferentiated cells. Current knowledge of surface CD profile and cell migration could be beneficial for regenerative cellular therapies.
Data provided are for informational purposes only. Although carefully collected, accuracy cannot be guaranteed. The impact factor represents a rough estimation of the journal's impact factor and does not reflect the actual current impact factor. Publisher conditions are provided by RoMEO. Differing provisions from the publisher's actual policy or licence agreement may be applicable.