Fig. 1

Recovering mES cells from cryopreservation with MEF or TM4 feeder. a At day 3, PCs were able to aggregate in group mES+TM4. Scale bar = 400 μm. Enlarged figure shows on the right. b Growth curves of mES cells in mES+MEF, mES+TM4, mES+Trans, and mES+Trans+TM4 groups in the first 5 days. Results were expressed as mean ± SD (n = 3 independent experiments). Asterisks indicate statistical significance calculated by one-way ANOVA method and labeled between groups mES+TM4 and mES+MEF at days 4 and 5. c Five pluripotency markers of cells were detected in groups mES+MEF, mES+TM4, and mES+Trans by qPCR at day 5. qPCR results were expressed relative to the expression in group mES+MEF (control group). Results were expressed as mean ± SD (n = 3 independent experiments). Asterisks indicate statistical significance of differences in the mean gene expression according to the control group (mES+MEF). d Transcriptional expression of lentiviral transduced factors in mES cells were detected in groups mES+MEF, mES+Trans, and mES+Trans+TM4. qPCR results show changes in introduced factors among different groups in gene expression relative to expression of β-actin in self-comparison. Results were expressed as mean ± SD (n = 3 independent experiments). Asterisks indicate statistical significance of differences in the mean gene expression between the indicated groups. e Transcriptional level of some early gonadogenesis markers in mES+TM4 were expressed relative to the expression in group mES+MEF (control group) at day 5. Results were expressed as mean ± SD (n = 3 independent experiments). Asterisks indicate statistical significance of differences in the mean gene expression according to the control group (mES+MEF). (*P value < 0.05, **P value < 0.01, ***P value < 0.001)