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Body mass index affects proliferation and osteogenic differentiation of human subcutaneous adipose tissue-derived stem cells.

Frazier TP, Gimble JM, Devay JW, Tucker HA, Chiu ES, Rowan BG - BMC Cell Biol. (2013)

Bottom Line: It was hypothesized that overweight BMI-ASCs would be more compromised in early adipogenic and osteogenic potential, and ability to form colonies in vitro.Alizarin red staining and RT-PCR for alkaline phosphatase demonstrated that elevated BMI resulted in compromised ASC mineralization of extracellular matrix and decreased alkaline phosphatase mRNA expression.These data demonstrate that elevated BMI resulted in reduced ASC proliferation, and potentially compromised osteogenic capacity in vitro; thus BMI is an important criterion to consider in selecting ASC donors for clinical applications.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Structural and Cellular Biology, Tulane University, New Orleans, LA, USA.

ABSTRACT

Background: Obesity is associated with a higher risk of developing cancer and co-morbidities that are part of the metabolic syndrome. Adipose tissue is recognized as an endocrine organ, as it affects a number of physiological functions, and contains adipose tissue-derived stem cells (ASCs). ASCs can differentiate into cells of multiple lineages, and as such are applicable to tissue engineering and regenerative medicine. Yet the question of whether ASC functionality is affected by the donor's body mass index (BMI) still exists.

Results: ASCs were isolated from patients having different BMIs (BMI-ASCs), within the ranges of 18.5-32.8. It was hypothesized that overweight BMI-ASCs would be more compromised in early adipogenic and osteogenic potential, and ability to form colonies in vitro. BMI was inversely correlated with ASC proliferation and colony forming potential as assessed by CyQUANT proliferation assay (fluorescence- based measurement of cellular DNA content), and colony forming assays. BMI was positively correlated with early time point (day 7) but not later time point (day 15) intracytoplasmic lipid accumulation as assessed by Oil-Red-O staining. Alizarin red staining and RT-PCR for alkaline phosphatase demonstrated that elevated BMI resulted in compromised ASC mineralization of extracellular matrix and decreased alkaline phosphatase mRNA expression.

Conclusions: These data demonstrate that elevated BMI resulted in reduced ASC proliferation, and potentially compromised osteogenic capacity in vitro; thus BMI is an important criterion to consider in selecting ASC donors for clinical applications.

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BMI did not significantly affect adipogenesis potential in vitro. ASC adipogenesis was induced using differentiation induction medium for three days. ASC cultures were then switched to maintenance medium until day 15. Lipid formation was assessed by percent incorporation of oil red-o (ORO) into monolayers cultured in adipocyte differentiation medium for a. 7 days, and b. 15 days. c. Representative micrographs of ORO staining in BMI-ASCs at day 15. d, e. Regression analyses of differentiation data that was analyzed at days 7 and 15 using least squares ordinary fit. R2 value reflected no significant correlation at late adipogenesis timepoints (e; Day 15); however, early adipogenesis (d; Day 7) R2 value reflected a correlation between BMI and adipogenic potential. Values are reported as N ± SE; * p > 0.05, ** p > 0.001.
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Figure 3: BMI did not significantly affect adipogenesis potential in vitro. ASC adipogenesis was induced using differentiation induction medium for three days. ASC cultures were then switched to maintenance medium until day 15. Lipid formation was assessed by percent incorporation of oil red-o (ORO) into monolayers cultured in adipocyte differentiation medium for a. 7 days, and b. 15 days. c. Representative micrographs of ORO staining in BMI-ASCs at day 15. d, e. Regression analyses of differentiation data that was analyzed at days 7 and 15 using least squares ordinary fit. R2 value reflected no significant correlation at late adipogenesis timepoints (e; Day 15); however, early adipogenesis (d; Day 7) R2 value reflected a correlation between BMI and adipogenic potential. Values are reported as N ± SE; * p > 0.05, ** p > 0.001.

Mentions: To investigate the effect of BMI on ASC differentiation, adipogenesis was induced by culturing ASCs in differentiation induction medium for three days followed by culture in maintenance medium until day 15. Differentiation was induced in both 3% and 10% serum. Lipid formation was assessed by percent intracytoplasmic incorporation of Oil Red-O (ORO) into monolayers at days 7 and 15 of adipogenesis. Oil-red-o staining at day 7 revealed a positive correlation between BMI and adipogenesis at early time points (as BMI increased, lipid accumulation increased; Figure 3a). Grouping of BMI-ASCs revealed that overweight BMI-ASCs had significantly higher Oil-Red-O staining (61.40 ± 5.139) compared to the lean BMI-ASCs (46.20 ± 2.70); p = 0.017 at day 7 (data not shown). Staining at day 15 revealed that BMI has no significant effect on ASC adipogenesis during late time points (Figure 3b). To further investigate the correlation between BMI and adipogenesis, nonlinear regression analyses were applied to adipogenesis data from days 7 and 15. R2 values reflected a correlation between BMI and adipogenesis at day 7 (R2 = 0.78; Figure 3c), and no correlation at day 15 (R2 = 0.57; Figure 3d). Representative photomicrographs of ORO staining in BMI-ASCs at day 15 are shown in Figure 3e.


Body mass index affects proliferation and osteogenic differentiation of human subcutaneous adipose tissue-derived stem cells.

Frazier TP, Gimble JM, Devay JW, Tucker HA, Chiu ES, Rowan BG - BMC Cell Biol. (2013)

BMI did not significantly affect adipogenesis potential in vitro. ASC adipogenesis was induced using differentiation induction medium for three days. ASC cultures were then switched to maintenance medium until day 15. Lipid formation was assessed by percent incorporation of oil red-o (ORO) into monolayers cultured in adipocyte differentiation medium for a. 7 days, and b. 15 days. c. Representative micrographs of ORO staining in BMI-ASCs at day 15. d, e. Regression analyses of differentiation data that was analyzed at days 7 and 15 using least squares ordinary fit. R2 value reflected no significant correlation at late adipogenesis timepoints (e; Day 15); however, early adipogenesis (d; Day 7) R2 value reflected a correlation between BMI and adipogenic potential. Values are reported as N ± SE; * p > 0.05, ** p > 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750383&req=5

Figure 3: BMI did not significantly affect adipogenesis potential in vitro. ASC adipogenesis was induced using differentiation induction medium for three days. ASC cultures were then switched to maintenance medium until day 15. Lipid formation was assessed by percent incorporation of oil red-o (ORO) into monolayers cultured in adipocyte differentiation medium for a. 7 days, and b. 15 days. c. Representative micrographs of ORO staining in BMI-ASCs at day 15. d, e. Regression analyses of differentiation data that was analyzed at days 7 and 15 using least squares ordinary fit. R2 value reflected no significant correlation at late adipogenesis timepoints (e; Day 15); however, early adipogenesis (d; Day 7) R2 value reflected a correlation between BMI and adipogenic potential. Values are reported as N ± SE; * p > 0.05, ** p > 0.001.
Mentions: To investigate the effect of BMI on ASC differentiation, adipogenesis was induced by culturing ASCs in differentiation induction medium for three days followed by culture in maintenance medium until day 15. Differentiation was induced in both 3% and 10% serum. Lipid formation was assessed by percent intracytoplasmic incorporation of Oil Red-O (ORO) into monolayers at days 7 and 15 of adipogenesis. Oil-red-o staining at day 7 revealed a positive correlation between BMI and adipogenesis at early time points (as BMI increased, lipid accumulation increased; Figure 3a). Grouping of BMI-ASCs revealed that overweight BMI-ASCs had significantly higher Oil-Red-O staining (61.40 ± 5.139) compared to the lean BMI-ASCs (46.20 ± 2.70); p = 0.017 at day 7 (data not shown). Staining at day 15 revealed that BMI has no significant effect on ASC adipogenesis during late time points (Figure 3b). To further investigate the correlation between BMI and adipogenesis, nonlinear regression analyses were applied to adipogenesis data from days 7 and 15. R2 values reflected a correlation between BMI and adipogenesis at day 7 (R2 = 0.78; Figure 3c), and no correlation at day 15 (R2 = 0.57; Figure 3d). Representative photomicrographs of ORO staining in BMI-ASCs at day 15 are shown in Figure 3e.

Bottom Line: It was hypothesized that overweight BMI-ASCs would be more compromised in early adipogenic and osteogenic potential, and ability to form colonies in vitro.Alizarin red staining and RT-PCR for alkaline phosphatase demonstrated that elevated BMI resulted in compromised ASC mineralization of extracellular matrix and decreased alkaline phosphatase mRNA expression.These data demonstrate that elevated BMI resulted in reduced ASC proliferation, and potentially compromised osteogenic capacity in vitro; thus BMI is an important criterion to consider in selecting ASC donors for clinical applications.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Structural and Cellular Biology, Tulane University, New Orleans, LA, USA.

ABSTRACT

Background: Obesity is associated with a higher risk of developing cancer and co-morbidities that are part of the metabolic syndrome. Adipose tissue is recognized as an endocrine organ, as it affects a number of physiological functions, and contains adipose tissue-derived stem cells (ASCs). ASCs can differentiate into cells of multiple lineages, and as such are applicable to tissue engineering and regenerative medicine. Yet the question of whether ASC functionality is affected by the donor's body mass index (BMI) still exists.

Results: ASCs were isolated from patients having different BMIs (BMI-ASCs), within the ranges of 18.5-32.8. It was hypothesized that overweight BMI-ASCs would be more compromised in early adipogenic and osteogenic potential, and ability to form colonies in vitro. BMI was inversely correlated with ASC proliferation and colony forming potential as assessed by CyQUANT proliferation assay (fluorescence- based measurement of cellular DNA content), and colony forming assays. BMI was positively correlated with early time point (day 7) but not later time point (day 15) intracytoplasmic lipid accumulation as assessed by Oil-Red-O staining. Alizarin red staining and RT-PCR for alkaline phosphatase demonstrated that elevated BMI resulted in compromised ASC mineralization of extracellular matrix and decreased alkaline phosphatase mRNA expression.

Conclusions: These data demonstrate that elevated BMI resulted in reduced ASC proliferation, and potentially compromised osteogenic capacity in vitro; thus BMI is an important criterion to consider in selecting ASC donors for clinical applications.

Show MeSH
Related in: MedlinePlus