Song F
PUMC Hospital, CAMS, Beijing.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1996 Aug;18(4):305-9.
Since the first report on the use of mutants lacking enzymes and HAT medium for hybrid selection (Littlefield, 1964), enormous similar techniques, at least, have been basically established. These techniques have opened new avenues to assigning genes, analysing gene regulation in eukaryotic cells and studying mechanisms which govern cell growth and differentiation during normal embryonic development, wound healing and tumor formation. But some problems still remain. For instance, such techniques need about one year to mutate both kinds of cells before fusion and selection, A simple technique of somatic cell hybridization without cell mutation has been developed, and by utilizing different appearances of cells before and after trypsinization, two kinds of cells may be fused and selected in some hours. Up-to-date six kinds of cells have been tested and found successful. Some hundreds of tests have been repeated, and modified, and the analyses by different methods have shown that the technique is both stable and reliable.
自从首次报道使用缺乏酶的突变体和HAT培养基进行杂交筛选以来(利特菲尔德,1964年),至少已经基本建立了大量类似技术。这些技术为基因定位、分析真核细胞中的基因调控以及研究在正常胚胎发育、伤口愈合和肿瘤形成过程中控制细胞生长和分化的机制开辟了新途径。但仍存在一些问题。例如,此类技术在融合和筛选之前需要大约一年时间使两种细胞发生突变。已经开发出一种无需细胞突变的体细胞杂交简单技术,通过利用胰蛋白酶消化前后细胞的不同外观,两种细胞可在数小时内进行融合和筛选。到目前为止,已经对六种细胞进行了测试并取得成功。已经重复并修改了数百次测试,不同方法的分析表明该技术既稳定又可靠。