Experimental study on the in vitro perfusion culture of human abdominal adipose tissue and its myogenic induction
10.3760/cma.j.issn.1009-4598.2018.07.018
- VernacularTitle: 人腹壁脂肪组织的体外灌注培养及其诱导成肌的实验研究
- Author:
Fangfei NIE
1
;
Yang AN
;
Qi LI
;
Jianxun MA
;
Zelian QIN
Author Information
1. Department of Plastic Surgery, Peking University Third Hospital, Beijing 100083, China
- Publication Type:Journal Article
- Keywords:
In vitro tissue culture;
perfusion culture;
adipose tissue;
myogenic induction
- From:
Chinese Journal of Plastic Surgery
2018;34(7):565-569
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the feasibility of in vitro perfusion culture of human adipose tissue and its induction into muscle tissue.
Methods:Human abdominal adipose tissue were cultured in vitro by perfusion culture. After 1, 3, 5 or 7 weeks, FAD/PI staining was used to detect the tissue vitality. Histological staining was used to observe the changes of its histomorphology. Protein expressions of myogenic molecules Myf-5 and myoD1 as well as muscle specific protein Desmin were measured by immunohistochemistry and Western blot assay.
Results:The adipose tissues cultured in myogenic induction media were still in the appearance of adipose tissue at 7 weeks. While in the basal medium without inducing, vascular pedicles shed after 7 weeks and could not continue to be cultured. FAD/PI staining showed that the tissue cultured in the induction media remained viable at 7 weeks, while the viability of the tissue in the basal culture medium decreased significantly at 5 weeks. Histologically, Myf-5, myoD1 and Desmin were all positively expressed in muscle tissues, while in adipose tissues, some mesenchymal and vascular endothelial cells expressed Myf-5 but not myoD1, and only separate vascular smooth muscle cells expressed Desmin. Interestingly, in adipose tissues cultured in myogenic induction medium, partial muscle-like tissue formed, evidenced as positive expression of Myf-5, myoD1 and Desmin. There was no muscle-like tissue formation in adipose tissue cultured in basal medium and the expression patterns were similar to that of the control group. Western blot results showed that the expression levels of Myf-5 and Desmin in muscle tissue were significantly higher than that of the other groups (P<0.05). The expression level of Myf-5 was slightly higher in inducing group than that in non-inducing group at the same time points, but the difference was not statistically significant. Similarly, the expression of Desmin was higher in inducing group than that in non-inducing group, yet there was statistically significant difference only in the first week. The protein expression of myoD1 was not detected in any group using Western blot.
Conclusions:The adipose tissue cultured in the perfusion bioreactor can survive for up to 7 weeks, and it can be partially induced into muscle-like tissue in the myogenic induction medium.
