Patterning pluripotent stem cells at a single cell level

Marina V. Pryzhkova, Greg M. Harris, Shuguo Ma, Ehsan Jabbarzadeh

Research output: Contribution to journalArticlepeer-review

Abstract

Studies of cell-extracellular matrix (ECM) interactions at a single cell level have drawn interest from scientists around the world. Subcellular ECM micropatterning techniques allow researchers to control cell shape, migration, and spindle orientation during mitosis potentially influencing the stem cell fate. Generally these studies have been limited to somatic cells rather than human pluripotent stem cells (hPSCs) which are capable of enormous differentiation potential. hPSCs require a defined ECM for attachment and express characteristic integrins mediating cell-substrate interactions. hPSCs also rely on cell-cell contacts for survival and to maintain self-renewal properties, but these circumstances also significantly limit hPSC observation at a single cell level. In addition, currently available methods for ECM micropatterning generally require a facility with trained personnel and intricate equipment to produce protein micropatterns. To overcome this problem, we have developed a protocol for vitronectin micropatterning using simple UV/ozone modification of polystyrene. Single hPSCs were able to attach and form characteristic stress fibers and focal adhesions similar to somatic cell types which demonstrate hPSC responsiveness to extracellular adhesive cues. Micropatterned hPSCs were able to be cultured for up to 48 hours while maintaining expression of pluripotency-associated transcription factor OCT4. Although further studies are necessary, the results of our investigation will potentially have a large impact on cell regenerative medicine and tissue engineering.

Original languageEnglish (US)
Pages (from-to)461-471
Number of pages11
JournalJournal of Biomaterials and Tissue Engineering
Volume3
Issue number4
DOIs
StatePublished - Aug 2013
Externally publishedYes

Keywords

  • Extracellular matrix
  • Micropatterning
  • Pluripotent stem cells
  • Polystyrene
  • Stress fibers
  • UV/Ozone
  • Vitronectin

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Medicine (miscellaneous)
  • Biomedical Engineering

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