BMC Cell Biology BioMed Central Research article Aging of mesenchymal stem cell in vitro

BackgroundA hot new topic in medical treatment is the use of mesenchymal stem cells (MSC) in therapy. The low frequency of this subpopulation of stem cells in bone marrow (BM) necessitates their in vitro expansion prior to clinical use. We evaluated the effect of long term culture on the senescence of these cells.ResultsThe mean long term culture was 118 days and the mean passage number was 9. The average number of PD decreased from 7.7 to 1.2 in the 10th passage. The mean telomere length decreased from 9.19 Kbp to 8.7 kbp in the 9th passage. Differentiation potential dropped from the 6th passage on. The culture's morphological abnormalities were typical of the Hayflick model of cellular aging.ConclusionWe believe that MSC enter senescence almost undetectably from the moment of in vitro culturing. Simultaneously these cells are losing their stem cell characteristics. Therefore, it is much better to consider them for cell and gene therapy early on.

[1]  J. Glowacki,et al.  Engraftment of a clonal bone marrow stromal cell line in vivo stimulates hematopoietic recovery from total body irradiation. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[2]  T. Brümmendorf,et al.  Telomere length and telomerase activity during expansion and differentiation of human mesenchymal stem cells and chondrocytes , 2003, Journal of Molecular Medicine.

[3]  P. Conget,et al.  Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells , 1999, Journal of cellular physiology.

[4]  R. Class,et al.  Propagation and senescence of human marrow stromal cells in culture: a simple colony‐forming assay identifies samples with the greatest potential to propagate and differentiate , 1999, British journal of haematology.

[5]  Robert A. Weinberg,et al.  Expression of TERT in early premalignant lesions and a subset of cells in normal tissues , 1998, Nature Genetics.

[6]  D J Prockop,et al.  Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[7]  S. Gerson,et al.  Rapid hematopoietic recovery after coinfusion of autologous-blood stem cells and culture-expanded marrow mesenchymal stem cells in advanced breast cancer patients receiving high-dose chemotherapy. , 2000, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[8]  Friedenstein Aj,et al.  Marrow microenvironment transfer by heterotopic transplantation of freshly isolated and cultured cells in porous sponges. , 1982 .

[9]  L. Hayflick THE LIMITED IN VITRO LIFETIME OF HUMAN DIPLOID CELL STRAINS. , 1965, Experimental cell research.

[10]  S. Gerson Mesenchymal stem cells: No longer second class marrow citizens , 1999, Nature Medicine.

[11]  Christian Clausen,et al.  Aging is associated with decreased maximal life span and accelerated senescence of bone marrow stromal cells. , 2003, Bone.

[12]  W. Hayes,et al.  Bone regeneration by implantation of purified, culture‐expanded human mesenchymal stem cells , 1998, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[13]  G. Saretzki,et al.  Mild hyperoxia shortens telomeres and inhibits proliferation of fibroblasts: a model for senescence? , 1995, Experimental cell research.

[14]  J. R. Smith,et al.  Aging of cells in culture. , 1984, International review of cytology.

[15]  I. Bellantuono,et al.  Study of Telomere Length Reveals Rapid Aging of Human Marrow Stromal Cells following In Vitro Expansion , 2004, Stem cells.

[16]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[17]  C. Harley,et al.  Evidence for a critical telomere length in senescent human fibroblasts. , 1995, Experimental cell research.

[18]  T. Jensen,et al.  Adult human mesenchymal stem cell as a target for neoplastic transformation , 2004, Oncogene.

[19]  R. Hoffman,et al.  Role of mesenchymal stem cells in hematopoietic stem cell transplantation , 2000, Current opinion in hematology.

[20]  H. Iba,et al.  Application of efficient and specific gene transfer systems and organ culture techniques for the elucidation of mechanisms of epithelial– mesenchymal interaction in the developing gut , 2000, Development, growth & differentiation.

[21]  A. Friedenstein,et al.  Marrow microenvironment transfer by heterotopic transplantation of freshly isolated and cultured cells in porous sponges. , 1982, Experimental hematology.

[22]  J. Hows,et al.  Adult bone marrow is a rich source of human mesenchymal ‘stem’ cells but umbilical cord and mobilized adult blood are not , 2003, British journal of haematology.

[23]  J. Cigudosa,et al.  Spontaneous human adult stem cell transformation. , 2005, Cancer research.