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Development of Recombinant Human Vitronectin Protein as Coating Matrix Reagent for Human ES or iPS C

Regenerative medicine offers the hope that cells for disease research and therapy might be created from readily available sources. To fulfill this promise, the available cells need to be converted into the desired cell types either in vivo or in vitro. Currently, there are two main approaches to accomplishing this goal: in vitro directed differentiation, which is used to push pluripotent stem cells, including embryonic stem cells or iPS cells, through steps similar to those occur during embryonic development; and reprogramming (also known as transdifferentiation), in which a differentiated cell is converted directly into the cell of interest without proceeding through a pluripotent intermediate. Both approaches require a unique cell culture system, in which, all chemicals or proteins are clearly defined for its quality and cost.

Stem cells are thought to reside within specific anatomical structure or “niches”, where they receive signals that maintain them in an undifferentiated state and induce their self –renewal as needed. Currently, feeder cells or Matrigel are two commonly used culture methods for sustaining ES cell culture. Lot variation in feeder cell production and undefined Matrigel mixture protein extracts from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells makes research data which derived from those two culture systems un-relevant to protocol standardization for clinical application development.

To develop a chemically defined ES (or iPS) culture system, recombinant human vitronectin protein was developed as a stand alone coating matrix reagent based on recent publication (1) and manufactured in E.coli using a proprietary “temperature-shift – Matrixbuffer” inclusion body refolding technology. In this report, various parameters related to different iPS generation technologies, such as retroviral delivery (3), intracellular TF protein delivery derived iPS cell method (PiPS) (4) and mRNA delivery based iPS system (5) were tested for further development of recombinant human vitronectin applications in ES or iPS cell culture.

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