Pluripotency Validation of Reprogrammed iPSCs

Alkaline phosphatase activity of the reprogrammed iPSC colonies was tested using an alkaline phosphatase detection kit (Millipore) according to the manufacturer's instructions. Immunocytochemistry was performed to confirm pluripotency marker expression (OCT4, 1 : 200, and TRA-1-81, 1 : 200, Abcam, Cambridge, UK). RT-PCR, karyotyping, and hematoxylin and eosin staining (HE staining) of teratoma tissues were performed as previously described [17 (link)].

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Chen I.C., Chang K.H., Chen Y.J., Chen Y.C., Lee-Chen G.J, & Chen C.M. (2019). Pueraria lobata and Daidzein Reduce Cytotoxicity by Enhancing Ubiquitin-Proteasome System Function in SCA3-iPSC-Derived Neurons. Oxidative Medicine and Cellular Longevity, 2019, 8130481.

Publication 2019

alkaline phosphatase detection kit TRA-1-81

Alkaline phosphatase Eosin Hematoxylin Immunocytochemistry Ipsc Oct4 Rt pcr Teratoma Tissues

Corresponding Organization : Chang Gung Memorial Hospital

Other organizations : National Taiwan Normal University

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Variable analysis

independent variables

Reprogramming of iPSC colonies

dependent variables

Alkaline phosphatase activity of the reprogrammed iPSC colonies
Pluripotency marker expression (OCT4 and TRA-1-81)
Teratoma formation

control variables

Manufacturer's instructions for alkaline phosphatase detection kit
Antibody dilutions for immunocytochemistry (OCT4 1:200, TRA-1-81 1:200)
Previously described methods for RT-PCR, karyotyping, and hematoxylin and eosin staining of teratoma tissues

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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