DERIVATION OF COMPLETELY CELL CULTURE-DERIVED MICE FROM EARLY-PASSAGE EMBRYONIC STEM-CELLS
DERIVATION OF COMPLETELY CELL CULTURE-DERIVED MICE FROM EARLY-PASSAGE EMBRYONIC STEM-CELLS
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DOI：
10.1073/pnas.90.18.8424
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发表时间：
1993-09-15
影响因子：
11.1
通讯作者：
RODER, JC
中科院分区：
文献类型：
--
作者：
NAGY, A;ROSSANT, J;RODER, JC
通过ES细胞四倍体胚胎聚集，测试了几种新生成的小鼠胚胎干（ES）细胞系在早期传代次数下产生完全ES细胞衍生小鼠的能力。一个线，指定为R1，产生活的后代，这是完全ES细胞衍生的同工酶分析和毛色判断。这些细胞培养衍生的动物是正常的、可存活的和可生育的。然而，长期的体外培养对R1的这种初始全能性产生了负面影响，在第14代之后，ES细胞衍生的新生儿在出生时死亡。然而，从第12代单细胞衍生的5个亚克隆之一（R1-S3）保留了原始的全能性，并产生了活的、完全ES细胞衍生的动物。试验时亚系的总体外培养时间相当于原系的第24代。完全有效的早期传代R1细胞和R1-S3亚克隆应该是非常有用的，不仅为ES细胞为基础的遗传操作，而且在定义最佳的体外培养条件，以保持初始全能性的ES细胞。
Several newly generated mouse embryonic stem (ES) cell lines were tested for their ability to produce completely ES cell-derived mice at early passage numbers by ES cell tetraploid embryo aggregation. One line, designated R1, produced live offspring which were completely ES cell-derived as judged by isoenzyme analysis and coat color. These cell culture-derived animals were normal, viable, and fertile. However, prolonged in vitro culture negatively affected this initial totipotency of R1, and after passage 14, ES cell-derived newborns died at birth. However, one of the five subclones (R1-S3) derived from single cells at passage 12 retained the original totipotency and gave rise to viable, completely ES cell-derived animals. The total in vitro culture time of the sublines at the time of testing was equivalent to passage 24 Of the original line. Fully potent early passage R1 cells and the R1-S3 subclone should be very useful not only for ES cell-based genetic manipulations but also in defining optimal in vitro culture conditions for retaining the initial totipotency of ES cells.