Induced pluripotent stem cells as a model for diabetes investigation

DOI: 10.1038/srep08597

Authors: J. Stepniewski; N. Kachamakova-Trojanowska; D. Ogrocki; M. Szopa; M. Matlok; M. Beilharz; G. Dyduch; M. T. Malecki; A. Jozkowicz; J. Dulak

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” unknown species cell and mouse PSC in “DMEM” for “1” days. For ECM, use “MEF feeder” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Ideally, “PSC” should express “alkaline phosphatase”, “HO-1”, “proteoglycan-4”, “angiotensinogen”, “Oct3/4A”, “NANOG” at the end of culture.

Step 2: mesoderm 1

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

culture “adherent” unknown species cell and mouse PSC in “unspecified medium”.

Ideally, “mesoderm” should be obtained at the end of culture.

Step 3: endothelial 1

day 2 ~ 6

culture “adherent” unknown species cell and mouse PSC in “alpha-MEM” for “4” days. For ECM, use “Collagen” at “0.05” “g/m2”. Medium should contain factors shown in the following table, and should be changed once in “4” days.

Step 4: endothelial 2

day 6 ~ 36

culture “adherent” unknown species cell and mouse PSC in “alpha-MEM” for “30” days. For ECM, use “FN1” at “0.02” “g/l”. Medium should contain factors shown in the following table, and should be changed once in “30” days.

Ideally, “EC” should express “VWF”, “NOS3”, “Flt-1”, “CD34”, “Tie-2” at the end of culture.