摘要
背景:脂肪干细胞是目前组织工程研究中的热点种子细胞,但由于其缺乏特异性的表面标记,故目前尚缺乏有效的纯化方法。通过纯化取材的脂肪来提高干细胞纯度是一种简单可行的方法。目的:分析在原代培养脂肪干细胞前,如何对取材的脂肪组织进行纯化。设计、时间及地点:观察对比动物实验,于2007-12/2008-03在上海市第六人民医院上海市实验医学动物中心完成。材料:选用4周龄SD大鼠,用于获得腹股沟处的脂肪组织。方法:解剖SD大鼠腹股沟处取材的脂肪组织,游离出其表面血管及内部的血管分支。剔除内部血管的同时将分布在血管周围的椭圆形结节组织一并切除。主要观察指标:对椭圆性结节行苏木精-伊红染色,显微镜下观察以确定其组织类型。分别对经过纯化处理和未纯化处理的脂肪组织行苏木精-伊红染色,显微镜下观察其组织结构上的差异。结果:①椭圆性小结节经苏木精-伊红染色后在显微镜下观察证实为淋巴组织。②纯化的脂肪组织结构一致,细胞成分单一;未纯化的脂肪组织结构紊乱,细胞种类多样、成分复杂。结论:脂肪干细胞原代培养所用的脂肪组织中含有多种组织成分,在剔除脂肪组织表面的血管、皮肤和肌肉组织的同时应一并剔除组织内的血管及其周围淋巴结。
BACKGROUND： Adipose tissue-derived stem cells receive a high attention in tissue engineering research. Adipose tissue-derived stem cells lack of specific surface marker, there is no effective pudfied method. Purified adipose is a simple method to elevate purify of stem cells. OBJECTIVE： To analyze how to purify adipose tissues before primary culture of adipose tissue-derived stem cells. DESIGN, TIME AND SETTING： The controlled animal experiment was performed at the Shanghai Animal Center of Experimental Medicine of Shanghai Sixth People＇s Hospital between December 2007 and March 2008. MATERIALS： Four-week old Sprague Dawley rats were used for obtaining adipose tissues from the inguinal groove. METHODS： Adipose tissues from rat inguinal groove were dissected to educe superficial blood vessel and blood vessel branches. Both blood vessel inside and elliptic nodal tissues surrounding blood vessels were excised. MAIN OUTCOME MEASURES： Stained elliptic nodal tissues stained by Hematoxylin-Eosin were observed with a microscope to make sure what kind of tissues they are. The purified adipose tissues and unpudfied adipose tissues were stained by Hematoxylin-Eosin. The differences in their tissue construction were observed using the microscope. RESULTS： Elliptic nodal tissues stained by Hematoxylin-Eosin were proved to be lymphatic tissues. The tissue construction of purified edipose tissues was pure, and the cellular component was simple. Conversely, the tissue construction of unpurified adipose tissues was complicated, and cells were various with complicated components. CONCLUSION： The component of adipose tissues used to primary cultured adipose tissue-derived stem cells is complicated. As resection of superficial blood vessel, skin and muscle tissues, blood vessel inside tissues and lymphatic tissues should also be excised.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第1期35-38,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
上海市科委临床重点课题基金(064119628)~~
