Generation of a human induced pluripotent stem cell (iPSC) line from a patient carrying a P33T mutation in the PDX1 gene

Xianming Wanga, b, i, Shen Chenc, Ingo Burtschera, b, Michael Sterra, b, Anja Hieronimusd, e, j, Fausto Machicaof, j, Harald Staigerd, g, j, Hans-Ulrich Häringd, e, j, Gabriele Ledererk, Thomas Meitingerh, k, Heiko Lickerta, b, i, j, ,
Source: Stem Cell Res
Publication Date: (2016)
Issue: 2: 273–276
Research Area:
Immunotherapy / Hematology
Stem Cells
Cells used in publication:
Fibroblast, dermal(NHDF-Ad), human adult
Species: human
Tissue Origin: dermal
Nucleofector® I/II/2b
4D-Nucleofector® X-Unit
Wang et al. generated an induced pluripotent stem cell (iPSC) line with a straight forward protocol. There are known genes where loss of function leads to pancreatic agenesis and associated with Diabetes Mellitus. The group selected biopsy material from one heterozygous PDX1 P33T carrier and isolated the fibroblasts from the skin. The experiments with nucleofection,-reprogramming -expansion resulted in positive hESC like colonies after 3 weeks. The karyotype showed no changes and the pluripotency has been shown by analysis of the stem cell markers and confirmed by in vivo experiments. This pluripotent iPSC will be very useful to study diabtes pathomechanisms. Another great example where the Yamanaka protocol in combination with Nucleofection worked well.
Homozygous loss-of-function mutations in the gene coding for the homeobox transcription factor PDX1 leads to pancreatic agenesis, whereas certain heterozygous point mutations are associated with Maturity-Onset Diabetes of the Young 4 (MODY4) and Type 2 Diabetes Mellitus (T2DM). To understand the pathomechanism of MODY4 and T2DM, we have generated iPSCs from a woman with a P33T heterozygous mutation in the transactivation domain of PDX1. The resulting PDX1 P33T iPSCs generated by episomal reprogramming are integration-free, have a normal karyotype and are pluripotent in vitro and in vivo. Taken together, this iPSC line will be useful to study diabetes pathomechanisms