Influence of Fetal Bovine Serum Protein Concentration in Adipose Tissue Extract on Adipogenic Differentiation of Adipose Derived Stem Cells

LI Ting, LI Yong, TIAN Wei-dong, GUO Wei-hua, YU Mei, LI Xiao-dong

Abstract

To observe whether fetal bovine serum (FBS) will affect the adipogenic ability of adipose derived stem cells (ADSCs) induced by adipose tissue extract (ATE), and to explore the effects of different FBS protein concentrations in ATE on adipogenic ability, cell proliferation and migration of ADSCs. Methods Rat ADSCs were cultured, passaged, and then subjected to osteogenic and neural induction. The adipose explants were cultured in culture medium with or without FBS, then ATE was collected to induce passage four (P4)-ADSCs, which were subsequently detected with oil red staining on the 7th day and the adipogenic ratio was calculated. Different concentrations of ATE without FBS (100 μg/mL, 250 μg/mL and 500 μg/mL) were used to induce P4-ADSCs before the adipogenic events, and the adipogenic ratio of each concentration group was observed. The effect of different protein concentrations on ADSCs proliferation and migration was also observed. Results Alizarin red staining and NF immunofluorescence staining were positive after the osteogenic and neural induction of cultured ADSCs. Either ATE with or without FBS was able to induce adipogenic differentiation of ADSCs on the 3rd day and there was no significant differences of adipogenic differentiation between ATE with FBS and without FBS on the 7th day. The adipogenic ratio of 500 μg/mL group was higher than that of 100 μg/mL group and 250 μg/mL group (P<0.05). After two days of induction, all the three different protein concentrations could inhibit cell proliferation, and different protein concentrations in ATE had no effect on the migration of ADSCs. Conclusion ATE obtained from culture medium without FBS has no effect on adipogenic capacity of ADSCs in a short period of time. The adipogenic ratio of ADSCs is associated with protein concentration in ATE.

 

 Keywords: Adipose tissue extract, Adipose derived stem cells, Concentration

 

Full Text:

PDF


References


Swan SH. Environmental phthalate exposure in relation to reproductive outcomes and other health endpoints in humans. Environ Res,2008; 108(2); 177-184.

Main KM.Mortensen GK.Kaleva MM. et al. Human breast milk contamination with phthalates and alterations of endogenous reproductive hormones in infants three months of age. Environ Health Perspect,2006; 114(2):270-276.

Yen TH, Lin-Tan DT. Lin JL. Food safety involving ingestion of foods and beverages preparedwith phthalate-plasticizer- containing clouding agents. J Formos Med Assoc, 2011; 110 (11):671-684.

Awal MA, Kurohmaru M, Andriana BB, el al. Mono-( 2- ethylhexyl) phthalate (MEHP) induces testicular alterations in male guinea pigs at prepubertal stage. Tissue Cell.2005;37(3); 167-175.

Borch J, Metzdorff SB, Vinggaard AM, el al. Mechanisms underlying the anti-androgenic effects of diethylhexyl phthalate in fetal rat testis. Toxicology,2006 ; 223(1-2); 144-155.

Sarraj MA, Escalona RM, Umbers A, et al. Fetal Testis Dysgenesis and Compromised Leydig Cell Function in Tgfbr3 (Betaglycan) Knockout Mice. Biol Reprod, 2010; 82 ( 1 ): 153- 162.

Chauvigne F, Menuet A, Lesne L, el al. Time- and dose- related effects of di-( 2-ethylhexyl) phthalate and its main metabolites on the function of the rat fetal testis in vitro. Environ Health Perspect ,2009; 117(4);515-521.

Bowman CJ, Tuener KJ,Sar M, et al. Altered gene expression during rat wolffian duct development following di ( n-butyl) phthalate exposure. Toxicol Sci,2005;86( 1); 161-174.

Akingbemi BT,Ge R, Klinefelter GR, et al. Phthalate-induced Leydig cell hyperplasia is associated with multiple endocrine disturbances. Proc Natl Acad Sci U S A, 2004; 101 ( 3) ; 775- 780.

Lovekamp ST, Davis BJ. Mechanisms of phthalate ester toxicity in the female reproductive system. Environ Health Perspect, 2003; 111 (2) ; 139-145.

Hallmark N. Walker M. Effects of monobutyl and di(n-butyl) phthalate in vitroon steroidogenesis and leydig cell aggregation in fetal testis explants from the rat: comparison with effects in vivoin the fetal rat and neonatal marmoset and in vitroin the human. Environ Health Perspect ,2007; 115(3) ;390-396.

Howdeshell KT, Furr J. Lambright CR, el at. Cumulative effects of dibutyl phthalate and diethylhexyl phthalate on male rat reproductive tract development: altered fetal steroid hormones and genes. Toxicol Sci, 2007 ; 99 ( 1): 190-202.

Erkekoglu P. Zeybekoglu P, Zeybek ND, et al. The effects of di(2-ethylhexyl) phthalate exposure and selenium nutrition on sertoli cell vimentin structure and germ-cell apoptosis in rat testis. Arch Environ Contam Toxicol.2012;62(3) :539-547.


Refbacks

  • There are currently no refbacks.