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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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Expression of alkaline phosphatase mRNA indicated that higher BMI was associated with reduced ASC osteogenesis capacity in vitro. ASC osteogenesis was induced using a cocktail medium for 9 days. RT-PCR was performed using primers and a probe to alkaline phosphatase for a measure of mRNA expression. ΔCT values were normalized to internal GAPDH. a. ungrouped averages of triplicates b. grouped averages of ‘lean’ (BMI <25 n = 5) and ‘overweight’ (BMI >25; n = 5) BMI-ASCs. c. Linear regression analyses of differentiation data that was analyzed at day 9 using least squares ordinary fit. R2 value reflected a strong correlation between BMI and osteogenesis. Values reported as N ± SE; * p = 0.017.
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Figure 5: Expression of alkaline phosphatase mRNA indicated that higher BMI was associated with reduced ASC osteogenesis capacity in vitro. ASC osteogenesis was induced using a cocktail medium for 9 days. RT-PCR was performed using primers and a probe to alkaline phosphatase for a measure of mRNA expression. ΔCT values were normalized to internal GAPDH. a. ungrouped averages of triplicates b. grouped averages of ‘lean’ (BMI <25 n = 5) and ‘overweight’ (BMI >25; n = 5) BMI-ASCs. c. Linear regression analyses of differentiation data that was analyzed at day 9 using least squares ordinary fit. R2 value reflected a strong correlation between BMI and osteogenesis. Values reported as N ± SE; * p = 0.017.

Mentions: To confirm the results of Figure 4 demonstrating a significant effect of BMI on ASC osteogenesis, alkaline phosphatase mRNA expression was measured in additional ASC cultures after induction of osteogenesis. Alkaline phosphatase mRNA results confirmed that when grouped, overweight BMI-ASCs were significantly more compromised in osteogenic differentiation potential (N ± SE: 0.71 ± 0.18) compared to the lean BMI-ASCs (N ± SE: 4.4 ± 0.21; p = 0.017,s Figure 5a,b). Nonlinear regression analyses were performed on osteogenesis RT-PCR data using least squares ordinary fit. The R2 value again reflected a strong inverse correlation (R2 = 0.89) between BMI and osteogenic potential (Figure 5c).


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)

Expression of alkaline phosphatase mRNA indicated that higher BMI was associated with reduced ASC osteogenesis capacity in vitro. ASC osteogenesis was induced using a cocktail medium for 9 days. RT-PCR was performed using primers and a probe to alkaline phosphatase for a measure of mRNA expression. ΔCT values were normalized to internal GAPDH. a. ungrouped averages of triplicates b. grouped averages of ‘lean’ (BMI <25 n = 5) and ‘overweight’ (BMI >25; n = 5) BMI-ASCs. c. Linear regression analyses of differentiation data that was analyzed at day 9 using least squares ordinary fit. R2 value reflected a strong correlation between BMI and osteogenesis. Values reported as N ± SE; * p = 0.017.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 5: Expression of alkaline phosphatase mRNA indicated that higher BMI was associated with reduced ASC osteogenesis capacity in vitro. ASC osteogenesis was induced using a cocktail medium for 9 days. RT-PCR was performed using primers and a probe to alkaline phosphatase for a measure of mRNA expression. ΔCT values were normalized to internal GAPDH. a. ungrouped averages of triplicates b. grouped averages of ‘lean’ (BMI <25 n = 5) and ‘overweight’ (BMI >25; n = 5) BMI-ASCs. c. Linear regression analyses of differentiation data that was analyzed at day 9 using least squares ordinary fit. R2 value reflected a strong correlation between BMI and osteogenesis. Values reported as N ± SE; * p = 0.017.
Mentions: To confirm the results of Figure 4 demonstrating a significant effect of BMI on ASC osteogenesis, alkaline phosphatase mRNA expression was measured in additional ASC cultures after induction of osteogenesis. Alkaline phosphatase mRNA results confirmed that when grouped, overweight BMI-ASCs were significantly more compromised in osteogenic differentiation potential (N ± SE: 0.71 ± 0.18) compared to the lean BMI-ASCs (N ± SE: 4.4 ± 0.21; p = 0.017,s Figure 5a,b). Nonlinear regression analyses were performed on osteogenesis RT-PCR data using least squares ordinary fit. The R2 value again reflected a strong inverse correlation (R2 = 0.89) between BMI and osteogenic potential (Figure 5c).

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