In the presence of LIF, both sunitinib and 2i were able to maintain the mESC morphology up to day 8 without passage (Supplementary information, Figure S3B).
Nearly 100% of the control, as well as Dex P1, D2 and D3 cells, were nestin positive, whereas no TuJ1- or GFAP-positive cells were detected at any passage (Supplementary Figure S2).
The results showed that the expression levels of α-SMA and CXCL12 were similar in all the CAFs at different passages, and E-cadherin was not expressed in any CAFs at different passages (Supplementary Figure 1B and C), which indicated that the fibroblats at low passages cultured in vitro retained the features of CAFs.
In addition, the analysis of multiple orthoxenograft passages revealed that this highly altered genome remained stable along successive xenograft passages (Supplementary Fig S4A and B).
Homogenous ciNPCs could be gradually achieved following passaging (Supplementary information, Figure S12).
We observed formation of branched structures over two passages (supplementary material Fig. S2A).
Confirmation of culture purity during ASC passaging (Supplementary Figure 1) eliminated the possibility of cytokine secretion by culture-contaminating hematopoietic cells.
Strikingly, T cell conditional medium was able to support serial expansion of MuSCs for at least 22 passages (Supplementary information, Figure S3D).
In marked contrast to these observations, the reprogramming of FA fibroblasts that were first transduced with the hTERT-LV and then subjected to gene editing generated 230 iPSC-like clones, most of which could be maintained after serial in vitro passages (Supplementary Fig S3).
The mosaic distribution of mtDNA depletion was also apparent at low cell passage number (Supplementary Material, Fig. S1), where occasional mtDNA-free (rho zero) cells were present (Supplementary Material, Fig. S1), interspersed with cells with apparently normal or near normal mtDNA levels.
