论文部分内容阅读
目的:研究肾脂肪囊内脂肪干细胞(ADSCs)的培养条件及生物学特性,为干细胞治疗肾脏疾病寻找更佳的细胞来源。方法:分别取大鼠肾脂肪囊及腹股沟区脂肪组织,分离培养ADSCs进行对比,细胞达80%融合时传代,观察细胞形态和增殖情况;采用MTT比色法测定细胞生长曲线;采用流式细胞术(FCM)检测细胞表面抗原的表达。结果:①相同质量肾脂肪囊和腹股沟区脂肪组织分离培养原代ADSCs显示,肾脂肪囊分离出ADSCs的数量显著超过腹股沟区脂肪组织;肾脂肪囊来源的ADSCs 5～6 d可达80%融合,而腹股沟区脂肪来源ADSCs则需要10～12 d。两者细胞均成梭形成纤维样细胞形态。②生长曲线分析显示肾脂肪囊ADSCs较腹股沟区ADSCs增殖迅速。③流式细胞术检测示两种干细胞的CD29抗原表达率均>99%,肾脂肪囊ADSCs CD44表达率为52.92%,腹股沟区ADSCs表达率为75.41%,内皮细胞标志抗原CD31均阴性。结论:与其他部位来源ADSCs相比,肾脂肪囊来源ADSCs具有分离量大,增殖迅速等特点,可在短时间内扩增到较大数量,可能成为治疗肾脏疾病的新的干细胞来源。
Objective: To study the culture conditions and biological characteristics of adipose-derived adipose derived stem cells (ADSCs) in kidney and to find a better source of stem cells for the treatment of kidney disease. Methods: ADSCs were isolated from rat kidney fat capsule and adipose tissue in the groin area, respectively. The cells were passaged at 80% confluency and the cell morphology and proliferation were observed. Cell growth curve was determined by MTT colorimetric assay. Flow cytometry The expression of cell surface antigen was detected by FCM. RESULTS: ① The primary ADSCs isolated from adipose tissue of the same quality in the kidney and groin area showed that the number of ADSCs isolated from the kidney adipose capsule significantly exceeded that of the inguinal area adipose tissue. ADSCs derived from the kidney adipose capsule reached 80% confluence in 5-6 days , While ADSCs in the groin area require 10-12 days. Both cells become shuttle forming fibroblast-like morphology. ② Growth curve analysis showed that ADSCs of kidney fat capsule proliferated more rapidly than ADSCs in the groin area. Flow cytometry showed that the expression of CD29 antigen in both stem cells was> 99%, the expression rate of CD44 in ADSCs was 52.92%, the expression rate of ADSCs in groin area was 75.41%, and CD31 of endothelial cell marker antigen was negative. CONCLUSION: Adipose-derived ADSCs derived from adipose-derived adipose tissue have the characteristics of large amount of separation and rapid proliferation compared with ADSCs derived from other parts. They can be expanded to a large number in a short time and may become new stem cell sources for treatment of kidney disease.