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. 2021 Dec 14;25(1):103525. doi: 10.1016/j.isci.2021.103525

Figure 2.

Figure 2

Efficient generation system of Nanog-GFP-positive miPSCs using SeVdp in feeder-free conditions

(A) Schematic of miPSC generation from Nanog-GFP MEF using SeVdp infection. On day 3 after infection, the medium was changed to serum replacement (SR)-based conditioned medium which was collected from the culture of the SL10 feeder cell line. In addition, Shield1 to stabilize KLF4 protein in the ProteoTuner Shield system and Blasticidin S to select the transduced cells were added on day 3 after infection until day 10 and day 7, respectively.

(B) A typical example of flow cytometric analysis of reprogrammed iPSCs in the conditioned medium or MEF medium using SeVdp on day 10 after infection.

(C) Flow cytometric analysis of the percentage of Nanog-GFP-positive cells after the infection of SeVdp carrying WT or L507A KLF4. Flow cytometric experiments were performed on days 5, 7, 10, 13, and 15 after the infection. Results are mean and SE, n = 5. p values calculated with t test were shown based on the comparison between WT and L507A KLF4 conditions.

(D) The number of Nanog-GFP-positive iPSC colonies were counted 13 days after infection of SeVdp to Nanog-GFP MEF from 10,000 cells. Results are mean and SE, n = 4. The p value calculated with the t test was shown based on the comparison between WT and L507A KLF4 conditions.