Bioorthogonal non-canonical amino acid tagging to track transplanted human induced pluripotent stem cell-specific proteome
DOI: 10.1186/s13287-024-03792-3
Authors: Divya Sridharan; Julie A. Dougherty; Uzair Ahmed; Shridhar K. Sanghvi; Syed Baseeruddin Alvi; Ki Ho Park; Helena Islam; Sue E. Knoblaugh; Harpreet Singh; Elizabeth D. Kirby; Mahmood Khan
Step 1: iPSC/ESC maintainance
day 0 ~ 1
culture “adherent” human PSC in “E8” for “1” days. “PSC” should be “upregulated” for “L274G”. For ECM, use “Cell Basement Membrane” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.
| concentration | unit | |
|---|---|---|
| Y27632 | 1.0e-5 | mol/l |
| polybrene | 0.006 | g/l |
| Puromycin | 0.001 | g/l |
| methionine | 1.2e-5 | mol/l |
| Anl | 1.3e-4 | mol/l |
Ideally, “PSC” should express “POU5F1”, “SSEA4”, “SOX2”, “TRA-1-60” at the end of culture.