Expansion of mesenchymal stem cells isolated from pediatric and adult donor bone marrow. J Cell Biochem

Department of Pediatrics, Regina Margherita Children's Hospital, University of Turin, Turin, Italy.
Journal of Cellular Biochemistry (Impact Factor: 3.26). 03/2006; 97(4):744-54. DOI: 10.1002/jcb.20681
Source: PubMed


The enormous plasticity of mesenchymal stem cells (MSCs) suggests an improvement of a standard protocol of isolation and ex vivo expansion for experimental and clinical use. We isolated and expanded MSCs from bone marrow (BM) of pediatric and young adult donors, to analyze the growth kinetic, immunophenotype, telomere length, karyotype during ex vivo expansion. Seventeen BM samples were collected from young adult donors and 8 from pediatric donors. MSCs isolated from two groups showed no morphological differences while their cell growth was strictly related to the donor's age. The MSCs isolated from pediatric donors reached a cumulative PD almost twice as high as MSCs isolated from young adult donors after 112 days (10.2 +/- 1.9 versus 5.5 +/- 3.7). Furthermore, we analyzed the modulation of antigen expression in the MSCs isolated from two groups until 10th passage (77 days) and there was no significant difference between the modulation of antigen expression. In particular, at the first passage, MSCs showed a low contamination of hemopoietic cells which became insignificant in the following passages. There was a high expression of CD90, CD29, CD44 and CD105 and variable and moderate expression of CD166 and CD106 at the start of MSC culture and at each passage during expansion. No chromosomal alteration or evidence of cellular senescence were observed in all analyzed samples. All these data suggest that MSCs can be isolated and expanded from most healthy donors, providing for an autologous source of stem cells.

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Available from: Simona Aschero, Sep 29, 2015
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    • "Furthermore , these cells had 78 chromosomes (76 autosomal and 2 sexual) (Langford et al. 1996; Mareschi et al. 2006). In this study, by assessing the first and eighth passages, no chromosomal disorders was observed in agreement with Mareschi et al. (2006) that evaluated the passages 2, 5, and 10 of human MSCs. In addition, karyotypes of the monkey BM-MSCs (Berman et al. 2010), placenta-derived MSCs in a woman (Semenov et al. 2010), endometrial MSCs in heifer (Mehrabani et al. 2014b), and Guinea pig fetal fibroblast cells (Mehrabani et al. 2014a) were normal and that of chromosomal analysis revealed no chromosomal abnormalities. "
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    ABSTRACT: Bone marrow is the most important source of mesenchymal stem cells (MSCs). Significant development in the comprehension of MSCs biology has opened the way to their clinical use. The object of this study was to establish a method for karyotyping of canine MSCs. We examined the effect of the increase in passage number on the chromosome and appearance of MSCs. A large 3-year-old mixed breed dog was used. Bone marrow (20 ml) was collected from the proximal humerus under general anesthesia and cultured and passaged in alpha minimal essential medium (αMEM) and fetal bovine serum (FBS). The canine bone marrow MSCs (BM-MSCs) were characterized by reverse transcription polymerase chain reaction (RT-PCR) and flask adherent fibroblast-like morphology. Karyotyping of cells of passages 1 and 8 was performed. The chromosome numbers were counted and evaluated to obtain the percentage of diploid cells. The best time for exposing BMMSCs with media containing colcemid was 10 h. These cells were positive for the BM-MSCs markers CD90 and negative for hematopoietic markers CD34 and CD45 by RT-PCR. Karyotyping of these cells was normal, and also fibroblast-like cells were observed in every passage.
    Comparative Clinical Pathology 07/2015; 24(1):1-6.. DOI:10.1007/s00580-015-2081-4 · 0.37 Impact Factor
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    • "Sources for stem cells vary, each of which have uses for certain diseases (2-4). Mesenchymal stem cells (MSCs) are one source for stem cells that are multipotent, non-hematopoietic and have the capability for self-renewal and differentiation (5,6). MSCs can be isolated from bone marrow (BM), cord blood, placenta, adipose tissue and liver (2,3,7,8) "
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    ABSTRACT: Stem cell therapy is a powerful technique for the treatment of a number of diseases. Stem cells are derived from different tissue sources, the most important of which are the bone marrow (BM), umbilical cord (UC) blood and liver. Human UC mesenchymal stem cells (hUC-MSCs) are multipotent, non-hematopoietic stem cells that have the ability to self-renew and differentiate into other cells and tissues such as osteoblasts, adipocytes and chondroblasts. In a number of reports, human and mouse models of disease have hUC-MSCs treatments. In this article, we review studies that pertain to the use of hUC-MSCs as treatment for diseases.
    Cell Journal 12/2014; 15(4):274-281. · 1.11 Impact Factor
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    • "In our study, passage 3 BM-MSCs obtained from different aged donors showed similar morphology in terms of their surface antigen characteristics. These results were in accordance with the study of Mareschi et al. (2006) where BM-MSCs of early adult donors (range of age 20–50) and pediatric donors (range of age 6–11) were compared until passage 10. The study by Huang et al. (2005) demonstrated that BM-MSCs of fetuses, 0–20-, 20–40-, and >40-year-old donors, had similar morphology and antigenic phenotype. "
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    ABSTRACT: Stem cell studies hold enormous potential for development of new therapies for tissue regeneration and repair. Bone marrow mesenchymal stem cells (BM-MSCs) can differentiate into a variety of non-hematopoietic tissues and contribute maintenance of healthy hematopoiesis by providing supportive cellular microenvironment into BM. Here, we investigated age-related differences in BM-MSCs by using attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy and FTIR imaging together with hierarchical clustering as a novel methods to clarify global alterations in the structure and function of macromolecules in characterized BM-MSCs of different aged donors. The results may contribute to identification of age-related new molecular marker(s) to determine the effects of donor age on MSCs. The spectral results reflected that there were significant increases in the concentration of saturated lipids, proteins, glycogen, and nucleic acids in children and adolescent group BM-MSCs when compared to the infants and early and mid adults. The concentration of mentioned macromolecules in adult (early and mid) BM-MSCs were significantly lower than the concentrations in the children and adolescents. These results were attributed to the increase in the proliferation activity in younger BM-MSCs. The distribution of macromolecules into the cells was shown as in the form of chemical maps by FTIR imaging, and the results are in agreement with the ATR-FTIR spectroscopy results. The cellular activity degree was determined by the thiazolyl blue tetrazolium bromide (MTT) proliferation assay to support ATR-FTIR spectroscopy results. BM-MSCs of five different age groups were discriminated by making the hierarchical cluster analysis where the spectral data according to alterations in structure and composition of macromolecules were considered.
    Journal of the American Aging Association 08/2014; 36(4):9691. DOI:10.1007/s11357-014-9691-7 · 3.39 Impact Factor
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