WHOLE ANIMAL-CELL SORTING OF DROSOPHILA EMBRYOS
WHOLE ANIMAL-CELL SORTING OF DROSOPHILA EMBRYOS
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DOI：
10.1126/science.1898782
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发表时间：
1991-01-04
期刊：
影响因子：
56.9
通讯作者：
NOLAN, GP
中科院分区：
文献类型：
--
作者：
KRASNOW, MA;CUMBERLEDGE, S;NOLAN, GP
原代培养细胞的使用受到无法净化大多数细胞的限制，尤其是从早期发育阶段开始的细胞。在整个动物细胞分选（WAC）中，源自具有LACZ转基因的动物的活细胞根据其β-半乳糖苷酶表达水平纯化，并具有荧光β-半乳糖苷酶底物和荧光激活的细胞分类。使用WACS，从早期的果蝇胚胎中纯化了表达构成基因的初期后室室细胞。神经元前体细胞也被纯化，它们分化为具有较高培养的神经元。由于有许多LACZ菌株，因此可能可以净化大多数类型的果蝇细胞。同样的方法也适用于其他生物体转化的生物。
Use of primary culture cells has been limited by the inability to purify most types of cells, particularly cells from early developmental stages. In whole animal cell sorting (WACS), live cells derived from animals harboring a lacZ transgene are purified according to their level of beta-galactosidase expression with a fluorogenic beta-galactosidase substrate and fluorescence-activated cell sorting. With WACS, incipient posterior compartment cells that express the engrailed gene were purified from early Drosophila embryos. Neuronal precursor cells were also purified, and they differentiated into neurons with high efficiency in culture. Because there are many lacZ strains, it may be possible to purify most types of Drosophila cells. The same approach is also applicable to other organisms for which germ-line transformation is possible.