In vitro perfusion culture of adipose tissue and its myogenic induction

Objective To investigate the myogenic possibility and mechanism of adipose tissue block induced by in vitro perfusion. Methods Myogenic inducing liquid was perfused through the vascular pedicle to cultrue the rat inguinal fat fascia flap. After 3 weeks, the tissue viability was detected by the fluorescein diacetate and propidium iodide (FAD/PI) staining. The changes of histomorphology was observed by histological staining and immunofluorescence staining was used to observe the expression of Desmin and myogenic differentiation factor 1(MyoD1). The expression levels of Desmin, myogenic factor 5(Myf-5)and MyoD1 were detected by Western blotting. Results FAD/PI staining showed that the adipose tissue cultured in the induction fluid was still viable at 3 weeks. Histological observation showed that the adipose tissues induced by the inducing medium had formed some muscle-like tissues locally. Immunofluorescence staining showed positive expression of Desmin and MyoD1. Protein quantitative results showed that compared with non-induced group (0.028 2±0.027 1), the expression of muscle specific protein Desmin in induced group (0.114 5±0.153 1) increased slightly, yet the difference had no statistical significance (t=0.622, P>0.05). The expression level of myogenic differentiation factor MyoD1 was higher in induced group (0.255 2±0.103 2) than non-induced group (0.023 9±0.024 6) and the difference was statistically significant (t=4.877, P<0.01). The reduction of Myf-5 in non-induced group was also inhibit in the induced group (the expression level of Myf-5 were 1 time in induced group and 0.13 times in non-induced group after the induction compared to the control fat tissue group), the difference had statistical significance (t=5.215, P<0.01). Conclusion The adipose tissues cultured in the perfusion bioreactor can survive at least 3 weeks in vitro, and they can be partially transformed into muscle-like tissues. Myogenic factor Myf-5 and MyoD1 are both involved in the myogenic process of adipose tissue in vitro. Key words: Adipose; Myogenic induction; Tissue culture; In vitro perfusion