A new method for isolating primary mesenchyme cells of the sea urchin embryo. Panning on wheat germ agglutinin-coated dishes.
A new method for isolating primary mesenchyme cells of the sea urchin embryo. Panning on wheat germ agglutinin-coated dishes.
复制标题
分离海胆胚胎原代间充质细胞的新方法。
DOI：
10.1016/0014-4827(87)90015-2
复制
发表时间：
1987
影响因子：
3.7
通讯作者：
McClay,DR
中科院分区：
文献类型：
--
作者：
Ettensohn,CA;McClay,DR
本文介绍了一种快速有效的分离海胆胚胎原代间充质细胞的方法。该程序包括三个简单的步骤：1.1.间充质囊胚在无钙人工海水中的解离.将所得的细胞悬浮液在已经用麦胚凝集素（WGA）包被的培养皿上孵育，PMC比其他细胞类型更牢固地粘附在WGA上。轻轻冲洗培养皿以去除松散附着的细胞，然后更剧烈地冲洗以去除PMC。pictus和Arbacia punctulata，并产生纯度为95-99%的PMC群体，如通过用异硫氰酸荧光素（FITC）-WGA和用特异性结合PMC的单克隆抗体染色的细胞的比例所确定的。PMCs的产量为4-5 × 106个细胞/100-mm培养皿，或1-2 × 107个PMCs/ml包装胚胎。这种方法的主要优点是，它可以快速，简单地进行，并产生纯的PMC群体。
This paper describes a rapid and efficient way to isolate primary mesenchyme cells (PMCs) of the sea urchin embryo. The procedure involves three simple steps:1.1. Dissociation of mesenchyme blastulae in calcium-free artificial seawater.2.2. Incubation of the resulting cell suspension on dishes that have been coated with wheat germ agglutinin (WGA), to which the PMCs adhere more firmly than do other cell types.3.3. Gentle rinsing of the dishes to remove loosely attached cells, followed by more vigorous rinsing to remove PMCs.This panning procedure has been applied to embryos of three species of sea urchins,Lytechinus variegatus, L. pictusandArbacia punctulata, and yields populations of PMCs that are 95–99% pure as determined by the proportion of cells that stain with fluorescein isothiocynate (FITC)-WGA and with a monoclonal antibody that binds specifically to PMCs. The yield of PMCs is 4–5 × 106cells/100-mm dish, or 1–2 × 107PMCs/ml of packed embryos. The principal advantages of this procedure are that it can be carried out rapidly and simply, and it yields pure populations of PMCs.