摘要
背景: 雌激素对脂肪干细胞向脂肪细胞诱导分化会有负向调节作用,但对长期深低温保存的肾脂肪囊来源脂肪间充质干细胞向脂肪细胞分化的效果尚未见报道。目的: 探讨雌激素对长期深低温保存的及新鲜提取的肾脂肪囊来源脂肪间充质干细胞成脂分化能力的影响。方法: 将长期深低温保存后复苏的及新鲜提取的第3代肾脂肪囊来源脂肪间充质干细胞分为4组进行成脂诱导分化,新鲜+雌激素组和冻存+雌激素组诱导时添加10-7mol/L雌激素,新鲜组和冻存组不添加。成脂诱导分化14d进行油红O染色及成脂量定量检测。结果与结论: 长期深低温保存后复苏的第3代肾脂肪囊来源脂肪间充质干细胞与新鲜提取的细胞形态和排列无差异;长期深低温保存后复苏的与新鲜提取的第3代肾脂肪囊来源脂肪间充质干细胞表面抗原分子CD29、CD44均呈阳性,CD31均呈阴性表达。成脂诱导后可观察到细胞内有脂滴形成,油红O染色呈阳性。在成脂诱导分化14d对各组进行成脂量比较,新鲜组与新鲜+雌激素组,冻存组与冻存+雌激素组的吸光度之间比较,差异有显著性意义,但新鲜组与冻存组之间差异无显著性意义。结果表明:小剂量雌激素可抑制长期深度低温保存后复苏的肾脂肪囊来源脂肪间充质干细胞的成脂分化;长期深低温保存后复苏的与新鲜提取的第3代肾脂肪囊来源脂肪间充质干细胞在成脂诱导方面无显著性差异。
METHODS： Passage 3 long-term cryopreserved and fresh adipose-derived mesenchymal stem cells from the kidney adipose capsule were divided into four groups, all of which were induced to adipogenic cells by induced fluid： fresh cells ＋ 10-7 mol/L estrogen, cryopreserved cells ＋ 10-7 mol/L estrogen, fresh cells and cryopreserved cells groups. Oil red O staining and adipogenic quantitative detection were performed at 14 days after induced adipogenic differentiation. RESULTS AND CONCLUSION： There were no differences in the morphology and arrangement between the cryopreserved and fresh cells. Both cryopreserved and fresh cells expressed CD29 and CD 44, but did not express CD31. Intracellular lipid droplets were observed after adipogenic differentiation by oil red O staining, and the cells were positive for oil red O staining. The adipogenic volume comparison among the four groups was detected on day 14 after adipogenic differentiation, and the absorbance values showed significant difference between the fresh cells and fresh cells ＋ estrogen groups, as well as between the cryopreserved cells andcryopreserved cells ＋ estrogen groups, but no difference between the fresh and cryopreserved cells groups. It is proved that low-dose estrogen can inhibit the adipogenic differentiation of long-term cryopreserved adipose-derived mesenchymal stem cells from the kidney adipose capsule; however, there is no significant difference between passage 3 long-term cryopreserved and fresh adipose-derived mesenchymal stem cells from the kidney adipose capsule in adipogenic differentiation.
出处
《中国组织工程研究》
CAS
CSCD
2013年第27期4998-5004,共7页
Chinese Journal of Tissue Engineering Research
基金
2011年辽宁省第一批次科学技术计划项目(2011225021)
肾脂肪囊来源脂肪干细胞向肾脏固有细胞分化及其在肾脂肪囊和肾组织中迁徙及归巢的实验研究~~
关键词
干细胞
脂肪干细胞
肾脂肪囊
脂肪间充质干细胞
长期深低温保存
复苏
成脂分化
雌激素
油红O染色
省级基金
干细胞图片文章
stem cells
adipose-derived stem cells
kidney adipose capsule
adipose-derived mesenchymal stem cells
long-term cryopreservation
resuscitation
adipogenic differentiation
estrogen
oil red 0 staining
provincialgrants-supported paper
stem cell photographs-containing paper
