C57BL／6J小鼠ES细胞系的建立及其特性分析

本文报道从C57BL/6J个鼠的囊胚中,建立了三个ES细胞系MESPU17,MESPU18,MESPU19。这些细胞的细胞学特征和强AKP反应,表明这三个细胞系具有干细胞的特征。这三个细胞系均为XY型,正常二倍体核型分别占70％、88％和59％。体外分化可形成简单类胚,体内分化可形成瘤块。与国际上通用的CCE和来自129/ter的ES细胞MESPU13相比,这三个细胞系的ES细胞较大;在体外培养时,生长较慢;细胞也较粘,进行显微注射时,很容易粘在注射针壁上。MESPU17,MESPU18进行了嵌合体制作,以BALB/c和昆明鼠的囊胚为受体,采用囊胚注射法未获嵌合体,但使用昆阴鼠的8细胞胚注射法和共培养法得到嵌合体。

ESTABLISHMENT AND CHARACTERISTICS OF ES CELL LINES DERIVED FROM C 57 BL/6 J MICE

Three embryonic stem cell lines, designated MESPU 17, MESPU 18 and MESPU 19 were establishcd from C57 BL/6 J blastocysts. The stem cell pattern of the new lines was demonstra-ted by cytological observations and strong AKP activity. All of the ES lines were XY type karyotype with normai diploid composition of 70%, 88% and 59% respectively. They were able to grow and differcntiate to form simple embryoid bodies in vitro and to form teratocar-cinoma tumors in vivo. Comparing these cells with CCE and MESPU 13 derived from 129/ter, the cells were comparatively large, the rate of growth of the cells was relatively slow in vitro, and the cells were considerably sticky and easier to adherent to microinjection pipet. No chimera of MESPU 17 or MESPU 18 was got by the method of the blastocyst injection of BALB/c or KMW (Kun Ming White), but by the msthod of injection of ES celis of MESPU 17 or MESPU 18 into 8-cell embryos of KMW and co-culture with KMW embryos, the chime-ras had been obtained.