Different culture media affect growth characteristics, surface marker distribution and chondrogenic differentiation of human bone marrow-derived mesenchymal stromal cells

Hagmann, Sébastien; Moradi, Babak; Frank, Sebastian; Dreher, Thomas; Kämmerer, Peer W.; Richter, Wiltrud; Gotterbarm, Tobias

doi:10.1186/1471-2474-14-223

Cited by 83 publications

(77 citation statements)

References 48 publications

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“…First of all, equine MSCs from different sources were Original Article shown to display differences in growth kinetics (36,46), differentiation potential (36,47), and gene expression (36), thus differences in surface marker expression could be expected as well. Furthermore, differences in the culture medium could influence the cell characteristics (48,49). In this study, to enable obtaining sufficient cell numbers from all MSC sources, DMEM supplemented with 20% FBS was used, instead of 10% which is more commonly used (31,33,39).…”

Section: Discussionmentioning

confidence: 99%

Comparative immunophenotyping of equine multipotent mesenchymal stromal cells: An approach toward a standardized definition

et al. 2014

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Horses are an approved large animal model for therapies of the musculoskeletal system. Especially for tendon disease where cell-based therapy is commonly used in equine patients, the translation of achieved results to human medicine would be a great accomplishment. Immunophenotyping of equine mesenchymal stromal cells (MSCs) remains the last obstacle to meet the criteria of the International Society for Cellular Therapy (ISCT) definition of human MSCs. Therefore, the surface antigen expression of CD 29, CD 44, CD 73, CD 90, CD 105, CD 14, CD 34, CD 45, CD 79a, and MHC II in equine MSCs from adipose tissue, bone marrow, umbilical cord blood, umbilical cord tissue, and tendon tissue was analyzed using flow cytometry. Isolated cells from the different sources and donors varied in their expression pattern of MSC-defining antigens. In particular, CD 90 and 105 showed most heterogeneity. However, cells from all samples were robustly positive for CD 29 and CD 44, while being mostly negative for CD 73 and the exclusion markers CD 14, CD 34, CD 45, CD 79a and MHC II. Furthermore, it was evident that enzymes used for cell detachment after in vitro-culture affected the detection of antigen expression. These results emphasize the need of standardization of MSC isolation, culturing, and harvesting techniques. As the equine MSCs did not meet all criteria the ISCT defined for human MSCs, further investigations for a better characterization of the cell type should be conducted. V C 2014 International Society for Advancement of Cytometry

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Section: Discussionmentioning

confidence: 99%

Comparative immunophenotyping of equine multipotent mesenchymal stromal cells: An approach toward a standardized definition

et al. 2014

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“…It is known that culture medium may affect expression of cell markers [27][28][29]. We have used serum-free RPMI 1640 as a culture medium which should have minimal impact on amnion in vitro to mimic in vivo conditions.…”

Section: Discussionmentioning

confidence: 99%

Growth factors and their receptors derived from human amniotic cells in vitro

Grzywocz

Pius-Sadowska

Kłos

et al. 2014

Folia Histochem Cytobiol.

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Abstract:In vitro studies have shown that amnion-produced growth factors participated in angiogenesis, re-epithelialization, and immunomodulation. The aim of our study was to investigate the growth factors and receptors produced by human amnion tissue and amniotic cells. Human amnions (hAM) were isolated, and amnion circles were dissected for in vitro analysis. Some amnion fragments were digested by the use of different methods to obtain two cell fractions, which were analysed for mesenchymal and epithelial cell markers. Amniotic circles and human amniotic cell fractions were cultured in a protein-free medium. Proteins secreted into the culture medium were analysed with a human growth factor antibody array. Conditioned culture media were added to human umbilical vein epithelial cells (HUVECs) to test for stimulation of migration (scratch test) and proliferation (Ki67 expression). Fraction 1 cells expressed both cytokeratin and mesenchymal cell markers which indicated that it was composed of a mixture of human amnion epithelial cells (hAECs) and mesenchymal stromal cells (hAMSCs). Fraction 2 cells mainly expressed cytokeratin and, therefore, were designed as hAECs. Secretion of proteins by the cultured cells increased with time. The hAM cultures secreted EGF-R, IGF, and IGFBP-2, -3 and -6; Cell Fraction 1 secreted NT-4, whereas Cell Fraction 2 secreted G-CSF, M-CSF, and PDGF. Conditioned media of hAM cultures stimulated HUVECs migration. We have showed for the first time that human amnions and amniotic cells secreted IGFBP-6, MCSF-R, PDGF-AB, FGF-6, IGFBP-4, NT-4, and VEGF-R3. We found that Cell Fraction 1, Cell Fraction 2, and the whole amnion secreted different proteins, possibly due to different proportions of amnion-derived cells and different cell-cell interactions. The hAM cell factors remained functional in vitro and induced intensified migration of HUVECs. The growth factors and receptors found in amnion or amniotic cell media might be used for regenerative medicine.

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“…Since tissue regeneration requires functional blood ves- In the absence of any universal and specific marker to define mesenchymal stem cells, their immunophenotyping relies on the use of combinations of both positive and negative markers. Note that MSCs profile may vary depending on the cell culture conditions [88] , or with their in situ localization [89] . Expression of the cell surface antigens CD73, CD90, CD105…”

Section: Mscs Are Perivascular Cellsmentioning

confidence: 99%

Brain mesenchymal stem cells: The other stem cells of the brain?

Appaix

Nissou

Sanden

et al. 2014

WJSC

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Multipotent mesenchymal stromal cells (MSC), have the potential to differentiate into cells of the mesenchymal lineage and have non-progenitor functions including immunomodulation. The demonstration that MSCs are perivascular cells found in almost all adult tissues raises fascinating perspectives on their role in tissue maintenance and repair. However, some controversies about the physiological role of the perivascular MSCs residing outside the bone marrow and on their therapeutic potential in regenerative medicine exist. In brain, perivascular MSCs like pericytes and adventitial cells, could constitute another stem cell population distinct to the neural stem cell pool. The demonstration of the neuronal potential of MSCs requires stringent criteria including morphological changes, the demonstration of neural biomarkers expression, electrophysiological recordings, and the absence of cell fusion. The recent finding that brain cancer stem cells can transdifferentiate into pericytes is another facet of the plasticity of these cells. It suggests that the perversion of the stem cell potential of pericytes might play an even unsuspected role in cancer formation and tumor progression.

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Different culture media affect growth characteristics, surface marker distribution and chondrogenic differentiation of human bone marrow-derived mesenchymal stromal cells

Cited by 83 publications

References 48 publications

Comparative immunophenotyping of equine multipotent mesenchymal stromal cells: An approach toward a standardized definition

Comparative immunophenotyping of equine multipotent mesenchymal stromal cells: An approach toward a standardized definition

Growth factors and their receptors derived from human amniotic cells in vitro

Brain mesenchymal stem cells: The other stem cells of the brain?

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