A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture

Sci Rep. 2015 Jun 11:5:11319. doi: 10.1038/srep11319.

Abstract

Factors limiting the adoption of iPSC technology include the cost of developing lines and the time period that it takes to characterize and bank them, particularly when integration free, feeder free, and Xeno-free components are used. In this manuscript we describe our optimization procedure that enables a single technician to make 20-40 lines at a time in a 24-96 well format in a reliable and reproducible fashion. Improvements spanned the entire workflow and included using RNA virus, reducing cytotoxicity of reagents, developing improved transfection and freezing efficiencies, modifying the manual colony picking steps, enhancing passaging efficiency and developing early criteria of success. These modifications allowed us to make more than two hundred well-characterized lines per year.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / economics
  • Cell Culture Techniques / methods*
  • Cell Line
  • Cellular Reprogramming Techniques / economics
  • Cellular Reprogramming Techniques / methods*
  • Cellular Reprogramming*
  • Culture Media / chemistry*
  • Humans
  • Induced Pluripotent Stem Cells*

Substances

  • Culture Media