The Induction Of Mouse IPS Cells By Controlled Folate Deprivation Method

Part â…  The cultivation of MEF under the condition of folate deprivation in vitroObjective To investigate the expression of reprogramming pluripotent genes of mouse embryonic fibroblast (MEF) cells, which were cultured in vitro under the null folate condition.Methods MEF cells were separately cultured in null folate medium and normal medium as control, compare the pluripotent gene(Oct4and Nanog) methylation state and pluripotency of gene expression of MEF cells between the two group cells.Results The Oct4promoter and Nanog promoter of MEF cells demethylate after cultured in folate-deficient media. And the expression level of Oct4and Nanog genes were improved by folate deprivation.Conclusion Folate deficiency could improve the Oct4and Nanog expression. Folate involved in cell reprogramming process. Part â…¡ Reprogramming of folate deprived mouse embryonic fibroblasts to pluripotent stem cells using4transcription factorsObjective To improve the induction efficiency of induced pluripotent stem (iPS) cells. Methods1) According to the results of the first part, under null folate condition cultured mouse embryo fibroblast (MEF) cells for3days as the experimental group; containing folic acid medium for3days as the control group.2) Using the classical4factors, the two group cells were reprogramming into iPS cells, and the iPS cells were identified, the efficiency were compared; then, the iPS cells from experimental group were karyotype ananlysis.Results The reprogramming efficiency of experimental group was0.05%, which was5-fold greater than control group (0.01%). Moreover, induced clones expressed pluripotent markers, and the iPS cells from experimental group show normal karyotype.Conclusion These findings demonstrated that folate deprivation method can improve the efficiency of mouse iPS cells. Part â…¢ Generation of mouse induced pluripotent stem cells by folate deprivated mouse embryonic fibroblasts with3transcription factorsObjective To reduce the oncogenicity of mouse pluripotent stem (iPS) cells.Methods1) under folate deprivated condition cultured mouse embryo fibroblast (MEF) cells for3days as the experimental group; containing folic acid medium for3days as the control group.2) The two groups cells were infected with retroviruses expressing Oct4, Sox2, and Klf4. Appraised them after the clones appeared.. Results The iPS clones were successfully obtained using3factors by folate deprivated MEF cells, reprogramming efficiency was0.01%, induced clones expressed pluripotent markers (AP, SSEA-1, and Oct4).Conclusion These findings illustrate folate play an important role in reprogramming, and folate deprivated could reduce the oncogenicity of iPS cells.