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目的分析体外大鼠脂肪干细胞(ADSCs)混合脂肪移植后的存活情况。方法无菌条件下切取大鼠腹股沟脂肪组织,消化、分离培养得到第3代ADSCs,行成骨诱导(茜素红染色)和成脂诱导(油红O染色)。用CM-Dil荧光标记第3代ADSCs,24只大鼠每只背部皮下3处分别植入1.5ml脂肪颗粒、1.5ml荧光标记的ADSCs(密度为5×107个细胞/ml)和0.9ml荧光标记的ADSCs+0.6ml脂肪颗粒。术后2周、1个月和3个月每次取出8只大鼠的移植物,石蜡切片HE染色观察病理变化,冰冻切片荧光显微镜下观察ADSCs定位。结果 ADSCs成骨诱导2周后茜素红染色阳性,成脂诱导3周后油红O染色阳性。ADSCs与脂肪颗粒混合移植能明显改善脂肪组织的病理学变化。结论体外分离培养的大鼠ADSCs具有成骨、成脂分化的潜能,能改善脂肪颗粒移植时的脂肪组织的液化吸收现象。
Objective To analyze the survival of rat adipose derived stem cells (ADSCs) mixed fat transplantation in vitro. Methods Under the condition of aseptic conditions, the rat inguinal adipose tissue was excised and digested. The 3rd passage ADSCs were obtained and osteogenic (alizarin red staining) and adipogenic induction (oil red O staining) were performed. The third generation of ADSCs were labeled with CM-Dil fluorescence. Twenty-four rats were implanted with 1.5 ml fat particles, 1.5 ml fluorescent labeled ADSCs (density 5 × 10 7 cells / ml) and 0.9 ml fluorescence Labeled ADSCs + 0.6 ml fat particles. After 2 weeks, 1 month and 3 months, 8 rats were removed for each time. The pathological changes of paraffin sections were observed by HE staining. The frozen sections were used to observe the localization of ADSCs under fluorescence microscope. Results ADSCs were positive for alizarin red staining 2 weeks after osteogenic induction, and oily red O staining was positive after 3 weeks of adipogenic induction. ADSCs mixed with fat particles can significantly improve the pathological changes of adipose tissue. Conclusion ADSCs cultured in vitro have the potential of osteogenic and adipogenic differentiation, which can improve the liquefaction and absorption of adipose tissue during the transplantation of fat particles.