快速纯化新生大鼠许旺细胞的新方法  

A Novel Approach to Rapid and High-efficiency Purification of Schwann Cells in Rats

作　　者：祝加学[1] 沈尊理[1] 秦金保[1] 张涛[2] 金羽青[1] 周广东[3] 

机构地区：[1]上海交通大学医学院附属第一人民医院整形外科,上海市200080 [2]上海交通大学医学院附属第九人民医院妇产科,上海市200011 [3]上海交通大学医学院附属第九人民医院整复外科,上海市200011

出　　处：《组织工程与重建外科杂志》2010年第3期141-144,共4页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：国家自然科学基金(30872630)

摘　　要：目的探讨一种简单高效的许旺细胞纯化方法。方法选用3～5d的SD乳鼠双侧坐骨神经,用DispaseⅡ短时消化去除神经外膜和束膜,用复合胶原酶NB4彻底消化、离心,获得原代细胞。细胞培养达到亚融合状态后,置于4℃冰箱约5min,收集首先脱壁的许旺细胞。最后用S100免疫荧光法和流式细胞仪法鉴定许旺细胞的纯度。结果通过酶消化法剥脱神经外膜和束膜,获得纯度85%以上的原代许旺细胞;经低温差速法进一步纯化,纯度可达99%。结论先用酶消化神经外膜和束膜,再用低温差速分离许旺细胞,能够获得高纯度的许旺细胞,是一种简便高效的纯化手段。Objective To explore a simple method for high-purity enrichment of Schwann cells （SCs）. Methods The sciatic nerve was dissected from both limbs of the neonatal Sprague-Dawley （SD） rat （3-5 days old）. The nerve was first digested by dispase Ⅱ to remove epineurium and perineurium, and then by collagenase NB4 to obtain primary SCs. The cells were centrifuged and cultured until confluence in a flask. Then the flask was placed in refrigerator （4 ℃） for 5 minutes. At such a low temperature, SCs mostly detached from the inner wall, while the other cells remained adherent. The SC purity was identified by S100 immunofluorescence double staining and flow cytometry. Results The purity was above 85% after enzyme digestion, and reached 99% after low-temperature treatment. Conclusion Enzyme digestion in conjunction with low temperature treatment is a simple and efficient way to obtain Schwann cells with high purity.

关 键 词：许旺氏细胞 纯化 低温 

分 类 号：Q813.11[生物学—生物工程]