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A link between the accumulation of DNA damage and loss of multi-potency of human mesenchymal stromal cells.

Alves H, Munoz-Najar U, De Wit J, Renard AJ, Hoeijmakers JH, Sedivy JM, Van Blitterswijk C, De Boer J - J. Cell. Mol. Med. (2010)

Bottom Line: Human mesenchymal stromal cells (hMSCs) represent an attractive cell source for clinic applications.Unfortunately, this leads to a significant decrease in their stemness.To identify the mechanism behind loss of multi-potency, hMSCs were expanded until replicative senescence and the concomitant molecular changes were characterized at regular intervals.

View Article: PubMed Central - PubMed

Affiliation: Department of Tissue Regeneration, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, Enschede, The Netherlands. hugoandrealves@gmail.com

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Long-term in vitro expansion of hMSCs leads to loss of multi-potency. (A) Growth profiles of hMSCs in vitro depicted by the number of population doublings per day plotted against culture period. (B) Phase-contrast photo of hMSCs at RPD 25. At RPD 25, most of the cells express SA-b-Galactosidase (black arrows). With increasing time of culture not only more cells were positive for SA- b-Galactosidase but also the staining was more intense (data not shown). (C) Effect of expansion in the expression of ALP, both in control and dexamethasone-treated cells. ALP expression was analysed by CDP-STAR base luminescence assay and corrected for the cell number. Error bars represent standard deviation. Statistical analysis was performed using one-way anova and Tukey post-test with a significance level of 0.05. * represents P < 0.01 to 0.05 and ***P < 0.001.
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fig01: Long-term in vitro expansion of hMSCs leads to loss of multi-potency. (A) Growth profiles of hMSCs in vitro depicted by the number of population doublings per day plotted against culture period. (B) Phase-contrast photo of hMSCs at RPD 25. At RPD 25, most of the cells express SA-b-Galactosidase (black arrows). With increasing time of culture not only more cells were positive for SA- b-Galactosidase but also the staining was more intense (data not shown). (C) Effect of expansion in the expression of ALP, both in control and dexamethasone-treated cells. ALP expression was analysed by CDP-STAR base luminescence assay and corrected for the cell number. Error bars represent standard deviation. Statistical analysis was performed using one-way anova and Tukey post-test with a significance level of 0.05. * represents P < 0.01 to 0.05 and ***P < 0.001.

Mentions: In order to investigate the differentiation capacity with respect to expansion, hMSCs from two donors were expanded until cessation of growth. As expected, hMSCs presented a continuous decrease in proliferation rate with increased time in culture (Fig. 1A). After an initial difference, both donors exhibited a similar level of PD/day until around 60 days in culture. At this point, the cells from donor 36 stopped proliferating while the cells from donor 24 displayed a temporary recovery followed by a permanent growth arrest. Although late passage cells did not proliferate, they were able to be maintained viable in culture for at least 3 more months before the experiment was ended. No spontaneous immortalization of hMSCs was ever observed and no clonal expansion was observed during this period of in vitro expansion.


A link between the accumulation of DNA damage and loss of multi-potency of human mesenchymal stromal cells.

Alves H, Munoz-Najar U, De Wit J, Renard AJ, Hoeijmakers JH, Sedivy JM, Van Blitterswijk C, De Boer J - J. Cell. Mol. Med. (2010)

Long-term in vitro expansion of hMSCs leads to loss of multi-potency. (A) Growth profiles of hMSCs in vitro depicted by the number of population doublings per day plotted against culture period. (B) Phase-contrast photo of hMSCs at RPD 25. At RPD 25, most of the cells express SA-b-Galactosidase (black arrows). With increasing time of culture not only more cells were positive for SA- b-Galactosidase but also the staining was more intense (data not shown). (C) Effect of expansion in the expression of ALP, both in control and dexamethasone-treated cells. ALP expression was analysed by CDP-STAR base luminescence assay and corrected for the cell number. Error bars represent standard deviation. Statistical analysis was performed using one-way anova and Tukey post-test with a significance level of 0.05. * represents P < 0.01 to 0.05 and ***P < 0.001.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3539748&req=5

fig01: Long-term in vitro expansion of hMSCs leads to loss of multi-potency. (A) Growth profiles of hMSCs in vitro depicted by the number of population doublings per day plotted against culture period. (B) Phase-contrast photo of hMSCs at RPD 25. At RPD 25, most of the cells express SA-b-Galactosidase (black arrows). With increasing time of culture not only more cells were positive for SA- b-Galactosidase but also the staining was more intense (data not shown). (C) Effect of expansion in the expression of ALP, both in control and dexamethasone-treated cells. ALP expression was analysed by CDP-STAR base luminescence assay and corrected for the cell number. Error bars represent standard deviation. Statistical analysis was performed using one-way anova and Tukey post-test with a significance level of 0.05. * represents P < 0.01 to 0.05 and ***P < 0.001.
Mentions: In order to investigate the differentiation capacity with respect to expansion, hMSCs from two donors were expanded until cessation of growth. As expected, hMSCs presented a continuous decrease in proliferation rate with increased time in culture (Fig. 1A). After an initial difference, both donors exhibited a similar level of PD/day until around 60 days in culture. At this point, the cells from donor 36 stopped proliferating while the cells from donor 24 displayed a temporary recovery followed by a permanent growth arrest. Although late passage cells did not proliferate, they were able to be maintained viable in culture for at least 3 more months before the experiment was ended. No spontaneous immortalization of hMSCs was ever observed and no clonal expansion was observed during this period of in vitro expansion.

Bottom Line: Human mesenchymal stromal cells (hMSCs) represent an attractive cell source for clinic applications.Unfortunately, this leads to a significant decrease in their stemness.To identify the mechanism behind loss of multi-potency, hMSCs were expanded until replicative senescence and the concomitant molecular changes were characterized at regular intervals.

View Article: PubMed Central - PubMed

Affiliation: Department of Tissue Regeneration, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, Enschede, The Netherlands. hugoandrealves@gmail.com

Show MeSH
Related in: MedlinePlus