[Comparison of myogenic differentiation ability of adipose-derived stem cells from different sites in rabbit].

ABSTRACT

OBJECTIVE: To compare the myogenic differentiation ability in vitro of rabbit adipose-derived stem cells (ADCSs) from different sites so as to provide ideal seed cells for repair and reconstruction of urinary tract. METHODS: Adipose tissues were obtained from the nape of the neck, post peritoneum, and vicinity of epididymis of a 4-month-old male New Zealand rabbit and ADSCs were harvested through collagenase digestion. ADSCs were purified by differential attachment method. The protein marker CD44 of rabbit ADSCs was used to identify the stem cells by immunocytochemistry, then the 5th generation of ADSCs were induced to differentiate into adipogenic, osteogenic, and myogenic cells. Multi-differentiation was confirmed by Oil red O staining, von Kossa staining, and RT-PCR. Myogenic differentiation abilities of ADSCs from 3 different sites were compared between the control group (L-DMEM medium containing 10%FBS) and the experimental group (myogenic medium) by RT-PCR method. RESULTS: ADSCs could be easily isolated from adipose tissues of the nape of the neck, post peritoneum, and vicinity of epididymis. ADSCs displayed a typical cobblestone morphology. Brown particles could be seen in ADSCs by CD44 immunocytochemistry staining. Oil red O staining showed red fat drops in ADSCs after 14 days of adipogenic culture. Black matrix could be seen in ADSCs by von Kossa staining after 28 days of osteogenic culture. RT-PCR detection showed moderate alpha-actin expression in the control group and strong alpha-actin expression in the experimental group after 42 days of myogenic culture. The growth rate of alpha-actin from the adipose tissue of post peritoneum (28.622% +/- 4.879%) was significantly lower (P < 0.05) than those from the adipose tissues of the nape of the neck (35.471% +/- 3.434%) and vicinity of epididymis (38.446% +/- 4.852%). CONCLUSION: The ADSCs from different sites show different myogenic differentiation abilities in vitro. ADSCs from the adipose tissues of the nape of the neck and vicinity of epididymis can be used as ideal seed cells for tissue engineering of lower urinary tract.