Transplantation of human induced pluripotent stem cell-derived neural crest cells for corneal endothelial regeneration
DOI: 10.1186/s13287-021-02267-z
Authors: Yajie Gong; Haoyun Duan; Xin Wang; Can Zhao; Wenjing Li; Chunxiao Dong; Zongyi Li; Qingjun Zhou
Step 1: iPSC/ESC maintainance
day 0 ~ 1 (duration missing in database; assumed 1 day)
culture “adherent” human PSC in “mTeSR”. For ECM, use “Matrigel” at “N/A” “N/A”.
Ideally, “PSC” should express “POU5F1”, “NANOG”, “SOX2” at the end of culture.
Step 2: Primitive streak 1
day 1 ~ 2
culture “unspecified method” human PSC in “DMEM/F12 medium” for “1” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.
| concentration | unit | |
|---|---|---|
| BSA | 2 | % |
| IGF1 | 2.0e-4 | g/l |
| Heregulinβ-1 | 1.0e-5 | g/l |
| FGF | 8.0e-6 | g/l |
| Ascorbic Acid | 0.05 | g/l |
| ITS | 1 | % |
| NEAA | 1 | % |
| 2-mercaptoethanol | 1.0e-4 | mol/l |
| GlutaMAX | 0.002 | mol/l |
| SB431542 | 2.0e-6 | mol/l |
| CHIR90021 | 1.0e-6 | mol/l |
| DMH1 | 1.0e-6 | mol/l |
| BMP4 | 1.5e-5 | g/l |
Ideally, “NCC” should express “P75”, “TFAP2A”, “TFAP2B”, “SOX10”, “SOX9”, “HNK-1”, “ZO-1”, “Na+/K+-ATPase”, “SNAIL2”, “ACTA2” at the end of culture.