ENRICHMENT OF SCHWANN-CELL CULTURES FROM NEONATAL RAT SCIATIC-NERVE BY DIFFERENTIAL ADHESION
ENRICHMENT OF SCHWANN-CELL CULTURES FROM NEONATAL RAT SCIATIC-NERVE BY DIFFERENTIAL ADHESION
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DOI：
10.1016/0006-8993(81)90375-9
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发表时间：
1981-01-01
期刊：
影响因子：
2.9
通讯作者：
PLEASURE, DE
中科院分区：
文献类型：
--
作者：
KREIDER, BQ;MESSING, A;PLEASURE, DE
建立了一种新的分离纯化新生大鼠坐骨神经雪旺细胞的方法。在酶促和机械解离后，使细胞消化物在间歇振荡下在聚赖氨酸包被的玻璃盖玻片上沉降30分钟。孵育18小时后，双极细胞占非粘附群体的> 95%。间接免疫荧光与细胞特异性标志物兔抗半乳糖苷和兔抗牛P-2碱性蛋白抗血清证实光镜鉴定这些双极细胞的雪旺细胞。兔抗人纤连蛋白特异性标记成纤维细胞，其占细胞群的< 5%，但不与许旺细胞结合。将差速贴壁法分离的雪旺细胞注射到家兔体内。当与培养的大鼠皮肤成纤维细胞吸收时，来自该兔的血清在体外18小时和5天后特异性地标记> 99%的双极和圆形细胞，并且还标记胎鼠背根神经节培养物中的雪旺细胞，但不标记成纤维细胞或神经元。
A novel method of Schwann cell purification from neonatal rat sciatic was developed using differential adhesion. After enzymatic and mechanical dissociation, the cell digest is allowed to settle on polylysine-coated glass coverslips for 30 min with intermittent shaking. After an 18 h incubation, bipolar cells comprise > 95% of the non-adherent population. Indirect immunofluorescence with the cell-specific markers rabbit anti-galactocerebroside and rabbit anti-bovine-P-2 basic protein antiserum confirmed light microscopic identification of these bipolar cells as Schwann cells. Rabbit anti-human fibronectin specifically labeled fibroblasts which comprised < 5% of the cell population, but did not bind to Schwann cells. Schwann cells isolated by differential adhesion were injected into a rabbit. When absorbed with cultured rat skin fibroblasts, serum from this rabbit specifically surface labeled > 99% of the bipolar and round cells after 18 h and 5 days in vitro and also labeled Schwann cells in fetal rat dorsal root ganglia cultures, but not fibroblasts or neurons.