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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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Related in: MedlinePlus

Higher BMI-ASCs exhibited compromised growth when exposed to low serum in vitro. a, b. CyQUANT was performed following ASC exposure to low serum (0-10% FBS supplementation) culture conditions for 48 hrs. Percentage serum and BMI both significantly affect ASC growth. c-f. Regression analysis of BMI-ASC growth when exposed to low serum in vitro. Nonlinear regression analysis performed using least squares ordinary fit. R2 values indicate a strong correlation between BMI, percent serum exposure, and ASC growth. Values are reported as N ± SE; * p > 0.05, ** p > 0.001, *** p > 0.0001.
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Figure 1: Higher BMI-ASCs exhibited compromised growth when exposed to low serum in vitro. a, b. CyQUANT was performed following ASC exposure to low serum (0-10% FBS supplementation) culture conditions for 48 hrs. Percentage serum and BMI both significantly affect ASC growth. c-f. Regression analysis of BMI-ASC growth when exposed to low serum in vitro. Nonlinear regression analysis performed using least squares ordinary fit. R2 values indicate a strong correlation between BMI, percent serum exposure, and ASC growth. Values are reported as N ± SE; * p > 0.05, ** p > 0.001, *** p > 0.0001.

Mentions: Cryopreserved ASCs isolated from lipoaspirates of women with different body mass indices (BMIs; Table 1), were cultured for up to 72 hours in ASC culture medium supplemented with 0 to 10% FBS. Cell growth was measured by MTT and CyQUANT cell proliferation assays. Growth data reflected an inverse relationship between BMI and ASC growth in vitro (Figure 1a,b). The largest effect was observed in 2% serum for 48 hrs (Figure 1a); however, growth was also compromised when ASCs were cultured 10% serum (Figure 1b). MTT data also revealed a time-dependent biphasic response in cell growth in which full recovery and maximum growth occurred at 72 hrs following a decline in higher BMI-ASC growth at 24 hrs and 48 hrs (data not shown). Non linear regression analyses using least fit ordinary squares supported the strong inverse relationship between both BMI (determination coefficient, R2 = 0.90; p < 0.05) and serum (R2 = 0.86; p < 0.05) on ASC growth (Figure 1c-f), where culture in the lowest percent serum (0%) reflected the strongest determination coefficient. Quadratic equations were used for nonlinear regression analyses, curve-fitting and subsequent R2 values (reported in Figure 1c-f).


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)

Higher BMI-ASCs exhibited compromised growth when exposed to low serum in vitro. a, b. CyQUANT was performed following ASC exposure to low serum (0-10% FBS supplementation) culture conditions for 48 hrs. Percentage serum and BMI both significantly affect ASC growth. c-f. Regression analysis of BMI-ASC growth when exposed to low serum in vitro. Nonlinear regression analysis performed using least squares ordinary fit. R2 values indicate a strong correlation between BMI, percent serum exposure, and ASC growth. Values are reported as N ± SE; * p > 0.05, ** p > 0.001, *** p > 0.0001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: Higher BMI-ASCs exhibited compromised growth when exposed to low serum in vitro. a, b. CyQUANT was performed following ASC exposure to low serum (0-10% FBS supplementation) culture conditions for 48 hrs. Percentage serum and BMI both significantly affect ASC growth. c-f. Regression analysis of BMI-ASC growth when exposed to low serum in vitro. Nonlinear regression analysis performed using least squares ordinary fit. R2 values indicate a strong correlation between BMI, percent serum exposure, and ASC growth. Values are reported as N ± SE; * p > 0.05, ** p > 0.001, *** p > 0.0001.
Mentions: Cryopreserved ASCs isolated from lipoaspirates of women with different body mass indices (BMIs; Table 1), were cultured for up to 72 hours in ASC culture medium supplemented with 0 to 10% FBS. Cell growth was measured by MTT and CyQUANT cell proliferation assays. Growth data reflected an inverse relationship between BMI and ASC growth in vitro (Figure 1a,b). The largest effect was observed in 2% serum for 48 hrs (Figure 1a); however, growth was also compromised when ASCs were cultured 10% serum (Figure 1b). MTT data also revealed a time-dependent biphasic response in cell growth in which full recovery and maximum growth occurred at 72 hrs following a decline in higher BMI-ASC growth at 24 hrs and 48 hrs (data not shown). Non linear regression analyses using least fit ordinary squares supported the strong inverse relationship between both BMI (determination coefficient, R2 = 0.90; p < 0.05) and serum (R2 = 0.86; p < 0.05) on ASC growth (Figure 1c-f), where culture in the lowest percent serum (0%) reflected the strongest determination coefficient. Quadratic equations were used for nonlinear regression analyses, curve-fitting and subsequent R2 values (reported in Figure 1c-f).

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