临床神经外科杂志2019，Vol.16Issue(1)：1-5,5.DOI:10.3969/j.issn.1672-7770.2019.01.001
新生大鼠原代小胶质细胞分离培养方法的改良
Modified method for cultivating primary microglia of neonatal rat
摘要
Abstract
Objective Isolation and culture of microglia in vitro has been widely used to explore new therapeutic strategies for various central nervous system diseases, but the existing protocols need a large amount of animal, higher cost and long culture time. Methods Combination of a unique combination of digestive reagents and shaking by hand were used to isolate and purify microglia. The purity of isolated cells was identified by expression of Iba-1 utilizing immunofluorescence technique and the viability of microglia was determined with trypan blue exclusion. Meantime, comparing the different states of microglia using the lipopolysaccharide test.Results The present modified methodology steadily yielded 1. 0 × 106 microglia per 60 mm dish with high purity (≥ 98％) and high survival rate (＞ 98％) within four days. Compared to the existing protocols, the animal dosage, total cost and culture time were reduced by 2, 3. 2 and 1. 5 times, respectively. Conclusion The improved method establishes a stable, simple and efficient cell culture model for studying the biological functions of microglia and elucidating the mechanisms of related CNS diseases.关键词
小胶质细胞/原代培养/新生大鼠Key words
microglia/primary culture/neonatal rat分类
医药卫生引用本文复制引用
钱凯,唐琳俊,王希,杨坤,张开鑫,钱腾达,马涛,石晶,李立新..新生大鼠原代小胶质细胞分离培养方法的改良[J].临床神经外科杂志,2019,16(1):1-5,5.基金项目
国家自然科学基金面上项目 (81171147) （81171147）
江苏省科教强卫工程医学重点人才研究基金 (ZDRCA2016010) （ZDRCA2016010）
江苏省兴卫工程医学重点人才基金 (RC201156) （RC201156）
