A human VE-cadherin-tdTomato and CD43-green fluorescent protein dual reporter cell line for study endothelial to hematopoietic transition

DOI: 10.1016/j.scr.2016.09.004

Authors: Ho Sun Jung; Gene Uenishi; Akhilesh Kumar; Mi Ae Park; Matt Raymond; Dustin Fink; Ethan McLeod; Igor Slukvin

Step 1: iPSC/ESC maintainance

day 0 ~ 1 (duration missing in database; assumed 1 day)

culture “adherent” human ESC in “E8”. For ECM, use “vitronectin” at “N/A” “N/A”. Medium should contain factors shown in the following table.

Ideally, “ESC” should express “POU5F1”, “SOX2”, “NANOG” at the end of culture.

Step 2: Primitive streak 1

day 1 ~ 2 (duration missing in database; assumed 1 day)

culture “adherent” unknown species cell in “unspecified medium”. For ECM, use “Collagen” at “N/A” “N/A”.

Step 3: Primitive streak 2

day 2 ~ 3 (duration missing in database; assumed 1 day)

culture “adherent” unknown species cell in “unspecified medium”. For ECM, use “Collagen” at “N/A” “N/A”.

Step 4: mesoderm 1

day 3 ~ 4 (duration missing in database; assumed 1 day)

culture “adherent” unknown species cell in “unspecified medium”. For ECM, use “Collagen” at “N/A” “N/A”.

Step 5: mesoderm 2

day 4 ~ 5 (duration missing in database; assumed 1 day)

culture “adherent” unknown species cell in “unspecified medium”. For ECM, use “Collagen” at “N/A” “N/A”.

Step 6: endothelial 1

day 5 ~ 13

culture “adherent” human ESC in “unspecified medium” for “8” days. For ECM, use “Collagen” at “N/A” “N/A”. Medium should be changed once in “8” days.

Ideally, “EC” should express “CDH5” at the end of culture.