Studies On Differential Expression Of Wnt Genes In Feeder Cells And Human Embryonic Stem Cells During Culture HES Cells In Vitro And RNAi For Wnt9a; Cloning And Priliminary Functional Study Of Novel Genes Cymg1 And Rcet1 Related To Spermatozoa Development

Objective: The aims of our research are â‘  to filter candidate Wnt genes which are important for maintenance of normal hESCs or resulting in karyotypic change through analysis of Wnt genes of differential expression in human and mouse feeder cells and hESCs during culture of normal or abnormal karyotype hESCs. â‘¡ to research the function of Wnt9a that RNAi for human Wnt9a were performed. â‘¢ to clone the spermatogenesis-related new genes expressed in mouse testes or reproductive tract and to study their function.Methods: â‘  differential expression analyses of Wnt genes in human and mouse feeder cells and hESCs during culture normal or abnormal karyotype hESCs were carried out to filter candidate Wnt genes which are important for maintenance of the normal hESCs or resulting in karyotypic change. Analyses of RT-PCR, in situ hybridization, immunohistochemistry and localization were performed to study the function of human Wnt9a. â‘¡ By synthesizing siRNAs, the pAVU6+27/siWnt9as expression vectors were constructed for human Wnt9a RNAi and then use in situ hybridization was performed to examine the effect of siRNAs for Wnt9a. MTT assay were carried out to analyze cell cycle and apoptosis at over expression and lower expression Wnt9a in MCF-7 cells. â‘¢ DDD and experiment methods were performed to clone new genes expressed in mouse testis or reproductive tract and to study their functions by RT-PCR, northern blotting, in situ hybridization, Immunohistochemistry, protein expression and cell location analysis.Results: â‘  Wnt7a was expressed in hEFCs and ICR mEFCs after culture normal karyotype hESCs, but not expressed in control and in hEFCs and ICR mEFCs after culturing abnormal karyotype hESCs with karyotypic change in chromosome NO. 1. Wnt3a was expressed in ICR and KUNMINGBAI mouse mEFCs after culturing normal karyotype hESCs, but not expressed in control and in mEFCs after culture abnormal karyotype hESCs. Wnt3 was expressed in ICR mEFCs and hEFCs after culture abnormal karyotype hESCs, not expressed in control, but...