Comparative characterization of mesenchymal bone marrow stromal cells at early and late stages of culturing

Biology Bulletin (Impact Factor: 0.25). 03/2008; 35(2):132-138. DOI: 10.1134/S1062359008020064


The mesenchymal stromal cell is a multipotent precursor of osteoblasts, adipocytes, and some other cell types. In this study,
a comparative analysis of cultured mesenchymal stromal cells from the rat bone marrow at the early and late stages of subculturing
has been performed using molecular genetic and cytological methods. The culture has undergone 11 passages during 140 days.
Upon long-term culturing, the mesenchymal stromal cells have proved to lose their potential for adipogenic differentiation
but preserve the potential for osteogenesis. Morphological characters typical of osteogenic differentiation can be observed
at the earlier stages of culturing (passages 1–4) but disappear at later stages (passages 9–11), despite mineralization of
the extracellular matrix and the expression of osteogenic differentiation markers. A comparative analysis of the proliferation
potential of stromal cells has shown that differences in the period of cell population doubling at the early and later stages
of culturing are insignificant. An almost complete arrest of cell growth has been observed in the middle of the culture period
(passages 5 and 6).

Download full-text


Available from: Arsen Mikaelyan, Apr 16, 2015
  • [Show abstract] [Hide abstract]
    ABSTRACT: First Page of the Article
    No preview · Conference Paper · Oct 1992
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: In recent years mesenchymal stem cells (MSCs) have generated a great deal of excitement as an attractive alternative to embryonic stem cells (ESCs) in cell-based regenerative medicine. In contrast to cells of embryonic origin, however, the clinical application of MSCs is heavily restricted by their finite ability of self-renewal, in which they resemble the rest of the somatic cells. Yet the mechanisms controlling MSC proliferation and senescence remain unclear. This review summarizes recent advances in our understanding of the factors affecting MSC expansion in vitro and discusses the pattern of their senescence with particular emphasis on the role of telomere shortening, activation of effectory pathways, and oxidative stress. The issues associated with MSC growth and senescence will be shown in the context of other somatic cells, and all of the parallels and disparities will be delineated precisely.
    Full-text · Article · May 2009 · Rejuvenation Research
  • [Show abstract] [Hide abstract]
    ABSTRACT: This study has aimed to repopulate 'primitive' cells from late-passage mesenchymal stem cells (MSCs) of poor multipotentiality and low cell proliferation rate, by simply altering plating density. Effects of low density culture compared t high density culture on late-passage bone marrow (BM)-derived MSCs and pluripotency markers of multipotentiality were investigated. Cell proliferation, gene expression, RNA interference and differentiation potential were assayed. We repopulated 'primitive' cells by replating late-passage MSCs at low density (17 cells/cm(2) ) regardless of donor age. Repopulated MSCs from low-density culture were smaller cells with spindle shaped morphology compared to MSCs from high-density culture. The latter had enhanced colony-forming ability, proliferation rate, and adipogenic and chondrogenic potential. Strong expression of osteogenic-related genes (Cbfa1, Dlx5, alkaline phosphatase and type Ι collagen) in late-passage MSCs was reduced by replating at low density, whereas expression of three pluripotency markers (Sox2, Nanog and Oct-4), Osterix and Msx2 reverted to levels of early-passage MSCs. Knockdown of Sox2 and Msx2 but not Nanog, using RNA interference, showed significant decrease in colony-forming ability. Specifically, knockdown of Sox2 significantly inhibited multipotentiality and cell proliferation. Our data suggest that plating density should be considered to be a critical factor for enrichment of 'primitive' cells from heterogeneous BM and that replicative senescence and multipotentiality of MSCs during in vitro expansion may be predominantly regulated through Sox2.
    No preview · Article · Oct 2011 · Cell Proliferation
Show more

Similar Publications