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通过一种使用生理条件的新方法分离的牛少突胶质细胞的特性。

Properties of bovine oligodendroglia isolated by a new procedure using physiologic conditions.

作者信息

Farooq M, Cammer W, Snyder D S, Raine C S, Norton W T

出版信息

J Neurochem. 1981 Feb;36(2):431-40. doi: 10.1111/j.1471-4159.1981.tb01611.x.

Abstract

A new class of procedures, previously shown to permit the isolation of pure oligodendroglia from whole rat cerebrum, has been applied with equal or greater success for the bulk isolation of this cell type from bovine white matter. Thus, the generality of this approach has been demonstrated. The bovine preparations have a purity of greater than 90% intact, phase-bright oligodendroglia and are obtained in a yield of 8 x 10(6) cells per gram of white matter. Within 1 day it is possible to obtain a preparation containing 60 mg of protein from a single cell type. These cells show a higher degree of ultrastructural preservation of all cytoplasmic constituents than previously obtained. The values for protein (33 pg/cell), DNA (5.4 pg/cell), and lipid (5-6 pg/cell) are very similar to those obtained with an earlier procedure. The cell lipids are rich in galactolipid, which comprises 20% of the total. The activity of the "myelin-specific" enzyme, 2',3'-cyclic nucleotide 3'-phosphohydrolase (EC 3.1.4.37), is 4.7 mumol/min/mg protein, similar to that obtained previously for isolated oligodendroglia and about 25-40% of that found in myelin. The activity of 5'-nucleotidase (EC 3.1.3.5) in the cells is about 10% of that in myelin or white matter.

摘要

一类新的程序,先前已证明可用于从整个大鼠大脑中分离出纯少突胶质细胞,现已同样成功或更成功地应用于从牛白质中大量分离这种细胞类型。因此,已证明了这种方法的通用性。牛的制备物中完整的、折光性强的少突胶质细胞纯度大于90%,每克白质的产量为8×10⁶个细胞。在1天内,从单一细胞类型中就有可能获得含60毫克蛋白质的制剂。这些细胞在所有细胞质成分的超微结构保存程度上比以前获得的细胞更高。蛋白质(33皮克/细胞)、DNA(5.4皮克/细胞)和脂质(5 - 6皮克/细胞)的值与早期方法获得的值非常相似。细胞脂质富含半乳糖脂,占总量的20%。“髓鞘特异性”酶2',3'-环核苷酸3'-磷酸水解酶(EC 3.1.4.37)的活性为4.7微摩尔/分钟/毫克蛋白质,与先前从分离的少突胶质细胞中获得的活性相似,约为髓鞘中该酶活性的25 - 40%。细胞中5'-核苷酸酶(EC 3.1.3.5)的活性约为髓鞘或白质中该酶活性的10%。

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