人脂肪来源基质细胞原代培养及成脂分化
发布时间：2018-12-20 10:17
【摘要】： 目的 建立人脂肪来源基质细胞(hADSCs)的原代培养方法,并进行脂肪诱导定向分化。探讨冻存后的ADSCs的特点和脂肪分化潜能,为临床应用提供实验室资料。 方法 取人腹部皮下脂肪,用酶消化法分离出单个核细胞,观察其生长情况。细胞冻存于液氮中6个月,复苏后观察其生长变化和脂肪分化潜能。细胞脂肪诱导以含地塞米松、1-甲基-3-异丁基-黄嘌呤、胰岛素的培养基作为脂肪定向诱导培养基。以油红O染色检测细胞内脂质沉积,观察细胞形态学变化和计数分化细胞数。 结果 原代培养的细胞形态均一,呈成纤维样,增殖力旺盛,可持续培养至15代。脂肪定向诱导培养后,第3天就可见细胞内核周有脂肪滴沉积。细胞形态的变化和油红O染色都证明这些细胞内有脂肪的沉积。诱导第9天细胞内出现大量脂质沉积,在17天左右达到高峰。2周以后,大约90%左右的细胞定向分化为成熟的脂肪细胞。冻存和新鲜的ADSCs生长特点相似,脂肪分化潜能没有差别(P0.5)。 结论 在成熟的脂肪组织中存在基质细胞,具有良好的自我增殖和脂肪分化潜能。冻存后的细胞仍能继续传代培养。并且能保持与冻存前相似的的脂肪分化潜能。ADSCs可用于脂肪组织工程种子细胞来源。
[Abstract]:Objective to establish a primary culture method of human adipose derived stromal cells (hADSCs) and to induce adipose differentiation. To explore the characteristics and adipose differentiation potential of frozen ADSCs, and to provide laboratory data for clinical application. Methods Mononuclear cells were isolated from human abdominal subcutaneous fat by enzyme digestion, and the growth of mononuclear cells was observed. The cells were frozen in liquid nitrogen for 6 months and their growth and adipose differentiation potential were observed after resuscitation. The adipose induction medium contained dexamethasone, 1-methyl-3-isobutyl xanthine and insulin. Oil red O staining was used to detect lipid deposition, morphological changes and number of differentiated cells were observed. Results the cells cultured in primary culture were homogeneous in morphology, fibroblast-like, strong in proliferation, and could be continuously cultured for 15 generations. Fat droplets were observed around the nucleus of the cells on the third day after adipose induction. Changes in cell morphology and oil red O staining all demonstrated fat deposition in these cells. On the 9th day after induction, a large number of lipid deposits appeared in the cells, and reached the peak at about 17 days. After 2 weeks, about 90% of the cells differentiated into mature adipocytes. The growth characteristics of frozen ADSCs were similar to those of fresh ADSCs, and there was no difference in adipose differentiation potential (P0. 5). Conclusion there are stromal cells in mature adipose tissue with good self-proliferation and adipose differentiation potential. Cryopreserved cells can continue to be subcultured. ADSCs can be used as a seed cell source for adipose tissue engineering.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329
本文编号：2387850
[Abstract]:Objective to establish a primary culture method of human adipose derived stromal cells (hADSCs) and to induce adipose differentiation. To explore the characteristics and adipose differentiation potential of frozen ADSCs, and to provide laboratory data for clinical application. Methods Mononuclear cells were isolated from human abdominal subcutaneous fat by enzyme digestion, and the growth of mononuclear cells was observed. The cells were frozen in liquid nitrogen for 6 months and their growth and adipose differentiation potential were observed after resuscitation. The adipose induction medium contained dexamethasone, 1-methyl-3-isobutyl xanthine and insulin. Oil red O staining was used to detect lipid deposition, morphological changes and number of differentiated cells were observed. Results the cells cultured in primary culture were homogeneous in morphology, fibroblast-like, strong in proliferation, and could be continuously cultured for 15 generations. Fat droplets were observed around the nucleus of the cells on the third day after adipose induction. Changes in cell morphology and oil red O staining all demonstrated fat deposition in these cells. On the 9th day after induction, a large number of lipid deposits appeared in the cells, and reached the peak at about 17 days. After 2 weeks, about 90% of the cells differentiated into mature adipocytes. The growth characteristics of frozen ADSCs were similar to those of fresh ADSCs, and there was no difference in adipose differentiation potential (P0. 5). Conclusion there are stromal cells in mature adipose tissue with good self-proliferation and adipose differentiation potential. Cryopreserved cells can continue to be subcultured. ADSCs can be used as a seed cell source for adipose tissue engineering.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329
【参考文献】
相关期刊论文 前1条
1 Raquel Taléns-Visconti;Ana Bonora;Ramiro Jover;Vicente Mirabet;Francisco Carbonell;José Vicente Castell;María José Gómez-Lechón;;Hepatogenic differentiation of human mesenchymal stem cells from adipose tissue in comparison with bone marrow mesenchymal stem cells[J];World Journal of Gastroenterology;2006年36期
，本文编号：2387850
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