ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2009, Vol. 40 ›› Issue (9): 1327-1332.doi:

• 遗传繁育 • Previous Articles     Next Articles

The Effects of Mouse ES Cell Conditioned Medium on Isolation and Cloning of Sheep Embryonic Stemlike Cells

BAI Changming 1,3，WANG Xinzhuang 1*，YAN Yingying 2，HE Xiaoe 1，ZHANG Jingfang 2

  

  1. 1. College of Husbandry and Animal Medicine Engineering，Henan Agricultural University， Zhengzhou 450002, China; 2. College of Veterinary Medicine, Northwest Agriculture and Forestry University, Yangling 712100, China; 3. Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China
  • Received:2008-09-05 Revised:1900-01-01 Online:2009-09-24 Published:2009-09-24
  • Contact: WANG Xinzhuang

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Abstract:

In order to evaluate the efficiency of mouse embryonic stem cells conditioned medium (ESCCM) and two kinds of feeder layers on selfrenewal of sheep embryonic stemlike cells, serumfree media was used to derive sheep embryonic stem (ES)like cells from embryos. Inner cell masses (ICM) were isolated and grown on the mouse embryonic fibroblasts (MEF) and sheep embryonic fibroblasts (SEF) as feeder layers respectively. The sheep ESlike cells were found to be easily obtained and remained undifferentiated for 10 passages in the new culture system with MEF, and 3 passages with SEF feeder layer. However, when cultured in medium without ESCCM, goat ESlike cells could not survive for more than 5 passages. Sheep ESlike cells isolated from ICMs had a normal karyotype and highly expressed alkaline phosphatase. Multiple differentiation potency of the ESlike cells was confirmed by expressing a novel ES cells specifically expressed transcription factor (Nanog) and forming embryoid bodies in suspension culture. These results suggest that mouse ES cells might secrete some factors which play important roles in promoting sheep ESlike cells′ selfrenewal. Additionally, the results also indicated that MEF is more suitable for the isolation and passage of sheep ESlike cells.