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目的:体外分离培养兔脂肪组织来源干细胞,初步探索其冻存复苏条件,为声带损伤修复重建的实验研究奠定基础。方法:分离兔腹股沟脂肪组织,胶原酶消化,低糖DMEM培养,倒置显微镜下观察其形态,MTT法检测其生长规律,定向诱导方法检测细胞的多向分化潜能,并进行冻存复苏。结果:分离培养的细胞具有很强的增殖能力和多向分化能力,在特定条件下可分化为脂肪细胞和成骨细胞,冻存复苏后的细胞仍能保持冻存前的生物特性。结论:成功分离培养出兔脂肪组织来源干细胞,其在液氮中能长期冻存且能复苏存活,可用于修复损伤的兔声带的实验研究。
OBJECTIVE: To isolate and culture rabbit adipose tissue-derived stem cells in vitro and to explore the conditions of their cryopreservation and resuscitation so as to lay the foundation for the experimental study of vocal cord injury repair and reconstruction. Methods: The rabbit groin adipose tissue, collagenase digestion and low glucose DMEM culture were isolated and observed under inverted microscope. The growth was observed by MTT assay. The multidrug differentiation potential of the cells was detected by directional induction method. Results: The isolated and cultured cells had strong proliferative ability and multidirectional differentiation ability, which could differentiate into adipocytes and osteoblasts under certain conditions. The cells after cryopreservation and resuscitation still maintained the biological characteristics before cryopreservation. CONCLUSION: Rabbit adipose tissue-derived stem cells are successfully isolated and cultured, which can be cryopreserved in liquid nitrogen for survival and can be resuscitated for survival. It can be used to repair damaged rabbit vocal cords.