Neverova M E, Kirsanova L A, Chentsov Iu S
Tsitologiia. 1985 Jul;27(7):785-91.
The modification of Prescott's (Prescott et al., 1972) method of enucleation in vitro was described. A special teflon chamber faciliatating the enucleation of monolayer cultured cells to produce cytoplasts and karyoplasts was constructed. Mouse L-cells were enucleated by exposing to cytochalasine B (10 gamma/ml) followed by centrifugation. The fraction of cells enucleated in the chamber was about 98%. The life time of cytoplasts in cultural medium after their enucleation was 48 hours (sometimes 56-72 hours) as tested by vital neutral red staining. The cytoplasts that survived were shown to accumulate large lysosomes, and the evidence of appearing ring-like fibrillar structures was provided using a simple technique of cytoskeleton observation under light microscope.
描述了对普雷斯科特(普雷斯科特等人,1972年)体外去核方法的改进。构建了一个特殊的聚四氟乙烯腔室,便于对单层培养细胞进行去核以产生胞质体和核质体。通过暴露于细胞松弛素B(10微克/毫升)然后离心对小鼠L细胞进行去核。在腔室中去核的细胞比例约为98%。通过活细胞中性红染色测试,胞质体去核后在培养基中的存活时间为48小时(有时为56 - 72小时)。存活的胞质体显示积累了大量溶酶体,并且使用一种在光学显微镜下观察细胞骨架的简单技术提供了出现环状纤维状结构的证据。