Adaptation of Human Testicular Niche Cells for Pluripotent Stem Cell and Testis Development Research

DOI: 10.1007/s13770-020-00240-0

Authors: Marina V. Pryzhkova; Philip W. Jordan

Step 1: iPSC/ESC maintainance

day 0 ~ 5

culture “adherent” human ESC in “E8” for “5” days. For ECM, use “Geltrex” at “N/A” “N/A”. Medium should be changed once in “5” days.

Step 2: Primitive streak 1

day 5 ~ 6

culture “spheroid” human ESC in “E8” for “1” days. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 3: Primitive streak 2

day 6 ~ 8

culture “spheroid” human ESC in “DMEM/F12” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 4: mesoderm 1

day 8 ~ 10

culture “spheroid” human ESC in “DMEM/F12” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Step 5: mesoderm 2

day 10 ~ 15

culture “device” human ESC in “DMEM/F12” for “5” days. Medium should contain factors shown in the following table, and should be changed once in “5” days.

Step 6: endothelial 1

day 15 ~ 16 (duration missing in database; assumed 1 day)

culture “device” human ESC in “DMEM/F12”. Medium should contain factors shown in the following table.

Ideally, “EC” should express “PECAM1” at the end of culture.

Step 7: endothelial 2

day 16 ~ 17 (duration missing in database; assumed 1 day)

culture “device” human ESC in “DMEM/F12”. Medium should contain factors shown in the following table.