Optimized Approaches for Generation of Integration-free iPSCs from Human Urine-Derived Cells with Small Molecules and Autologous Feeder

Stem Cell Reports. 2016 May 10;6(5):717-728. doi: 10.1016/j.stemcr.2016.04.001. Epub 2016 Apr 28.

Abstract

Generation of induced pluripotent stem cells (iPSCs) from human urine-derived cells (hUCs) provides a convenient and non-invasive way to obtain patient-specific iPSCs. However, many isolated hUCs exhibit very poor proliferation and are difficult to reprogram. In this study, we optimized reprogramming approaches for hUCs with very poor proliferation. We report here that a compound cocktail containing cyclic pifithrin-a (a P53 inhibitor), A-83-01, CHIR99021, thiazovivin, NaB, and PD0325901 significantly improves the reprogramming efficiency (170-fold more) for hUCs. In addition, we showed that replacement of Matrigel with autologous hUC feeders can overcome the reprogramming failure due to the massive cell death that occurs during delivery of reprogramming factors. In summary, we describe improved approaches to enable iPSC generation from hUCs that were otherwise difficult to reprogram, a valuable asset for banking patient-specific iPSCs.

Keywords: CPFT-a; P53; autologous UC feeders; hUCs; iPSCs; small molecules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Benzamides / pharmacology
  • Benzothiazoles / pharmacology
  • Cell Differentiation / drug effects*
  • Cell Proliferation / drug effects*
  • Cellular Reprogramming / drug effects*
  • Diphenylamine / analogs & derivatives
  • Diphenylamine / pharmacology
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / drug effects*
  • Pyrazoles / pharmacology
  • Pyridines / pharmacology
  • Pyrimidines / pharmacology
  • Thiazoles / pharmacology
  • Thiosemicarbazones / pharmacology
  • Toluene / analogs & derivatives
  • Toluene / pharmacology
  • Urine / cytology

Substances

  • A-83-01
  • Benzamides
  • Benzothiazoles
  • Chir 99021
  • PD 0325901
  • Pyrazoles
  • Pyridines
  • Pyrimidines
  • Thiazoles
  • Thiosemicarbazones
  • thiazovivin
  • Toluene
  • Diphenylamine
  • pifithrin