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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 negatively correlated with ASC colony-forming potential in vitro. ASCs were plated at a concentration of 100 cells/mL in 6 well plates and cultured for 14 days. The number of colonies per plate divided by the cells plated x 100 was determined as the “% CFU”. a. Ungrouped BMI-ASC colony-forming potential, and b. grouped BMI-ASC CFU potential. c. Visualization of colonies formed in 100 cm2 dishes. Values reported as N ± SE; * p > 0.05.
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Figure 2: BMI negatively correlated with ASC colony-forming potential in vitro. ASCs were plated at a concentration of 100 cells/mL in 6 well plates and cultured for 14 days. The number of colonies per plate divided by the cells plated x 100 was determined as the “% CFU”. a. Ungrouped BMI-ASC colony-forming potential, and b. grouped BMI-ASC CFU potential. c. Visualization of colonies formed in 100 cm2 dishes. Values reported as N ± SE; * p > 0.05.

Mentions: To further examine differences in BMI-ASC growth, colony-forming unit (CFU) assays were performed on the BMI-ASC donors (Table 1). These donors were grouped as follows: lean (BMI <25; mean BMI 22.2 ± 1.79, N = 5), and overweight (BMI >30; mean BMI 30.3 ± 2.17, N = 5). When grouped, the lean BMI-ASCs formed a significantly higher percentage of colonies (34.94 ± 1.46) compared to the overweight BMI-ASCs (28.26 ± 1.78); p < 0.05 (Figure 2b). Representative photomicrographs of CFUs are shown in Figure 2c. Annexin-V/PI staining and fluorescence-activated cell sorting (FACS) was used to determine whether the compromised growth in higher BMI-ASCs was accompanied by elevated apoptosis. There was no significant difference in apoptosis between lean and overweight BMI-ASCs at 24 and 48 hrs of culture with 2% or 10% FBS (data not shown). The percent early apoptotic cells did not exceed 5.49 ± 1.86%, and the percent late apoptotic cells did not exceed 4.12 ± 0.23% (mean ± SD).


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 negatively correlated with ASC colony-forming potential in vitro. ASCs were plated at a concentration of 100 cells/mL in 6 well plates and cultured for 14 days. The number of colonies per plate divided by the cells plated x 100 was determined as the “% CFU”. a. Ungrouped BMI-ASC colony-forming potential, and b. grouped BMI-ASC CFU potential. c. Visualization of colonies formed in 100 cm2 dishes. Values reported as N ± SE; * p > 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: BMI negatively correlated with ASC colony-forming potential in vitro. ASCs were plated at a concentration of 100 cells/mL in 6 well plates and cultured for 14 days. The number of colonies per plate divided by the cells plated x 100 was determined as the “% CFU”. a. Ungrouped BMI-ASC colony-forming potential, and b. grouped BMI-ASC CFU potential. c. Visualization of colonies formed in 100 cm2 dishes. Values reported as N ± SE; * p > 0.05.
Mentions: To further examine differences in BMI-ASC growth, colony-forming unit (CFU) assays were performed on the BMI-ASC donors (Table 1). These donors were grouped as follows: lean (BMI <25; mean BMI 22.2 ± 1.79, N = 5), and overweight (BMI >30; mean BMI 30.3 ± 2.17, N = 5). When grouped, the lean BMI-ASCs formed a significantly higher percentage of colonies (34.94 ± 1.46) compared to the overweight BMI-ASCs (28.26 ± 1.78); p < 0.05 (Figure 2b). Representative photomicrographs of CFUs are shown in Figure 2c. Annexin-V/PI staining and fluorescence-activated cell sorting (FACS) was used to determine whether the compromised growth in higher BMI-ASCs was accompanied by elevated apoptosis. There was no significant difference in apoptosis between lean and overweight BMI-ASCs at 24 and 48 hrs of culture with 2% or 10% FBS (data not shown). The percent early apoptotic cells did not exceed 5.49 ± 1.86%, and the percent late apoptotic cells did not exceed 4.12 ± 0.23% (mean ± SD).

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