[ @entity1 XXXX cell lines preliminarily established in China].

@entity1 @entity2085 @entity1390 @entity11823 @entity44 @entity19 @entity1138 @entity59908 @entity466
OBJECTIVE: To establish @entity2085 ( @entity2085 ) cell lines from @entity1 fertilized ovum in China. METHODS: Fourteen @entity1 oocytes obtained from a volunteer were cultured in Earlg's medium and fertilized in vitro. Four of the morulae formed from the fertilized ovi were frozen and five continued to develop into blastocysts. The pellucid zones of the 5 blastocysts were removed and their inner cell masses (ICM) were taken out and inoculated onto the feeder layer of @entity19 embryonic fibroblasts inactivated with @entity1138 . The cell clones formed from the three surviving ICM were selected and dispersed mechanically and the cells were inoculated onto new feeder layers to produce more cell clones. When multiple clones were produced, they were dissociated with dispase into smaller cell masses which were inoculated onto new feeder layer again for subculturing. During different periods of cell culture, alkaline phosphatase activity (AKP) staining, stem cell surface marker test, and karyotyping were conducted. The suspension of stem cells subcultured for 5 months were inoculated to the legs of @entity466 ( @entity466 ) @entity19 subcutaneously to observe the @entity44 formation. RESULTS: Out of the 9 fertilized oocytes, 5 developed into blastocysts. The ICM of three from the five blastocysts survived and developed into embronic stem cell lines, named @entity59908 , @entity1390 , and CHE3 respectively. These stem cells remained undifferentiated after subculturing for 7 months. The stem cell line CHE3 had been grown for 40 passages in vitro, @entity59908 for 36 passages, and @entity1390 for 32 passages. AKP staining was positive at the passages 5, 15, and 30 for the 3 cell lines. The surface markers of @entity1 stem cell, SSEA-4, SSEA-3, TRA-1-60, and GCTM-2 were positive at passage 25, 30, 40 for CHE3, and at the passages 21, 22 and 30 for @entity59908 and @entity1390 . But all cell lines did not express @entity11823 , a marker for @entity19 @entity2085 cells. All cell lines stably retained a normal karyotype. After routine passage by 5 months, @entity1 @entity2085 cells from 3 cell lines were injected subcutaneously into @entity466 @entity19 . Six weeks after inoculation of the 3 cell lines teratomas were formed in all @entity19 . Histological examination revealed that they contained various tissues derived from all three embryonic germ layers. CONCLUSION: @entity1 @entity2085 cell lines were successfully established in China.
