title: Engineered polymer-media interfaces for the long-term self-renewal of human embryonic stem cells.
abstract: We have developed a synthetic polymer interface for the long-term self-renewal of human embryonic stem cells (hESCs) in defined media. We successfully cultured hESCs on hydrogel interfaces of aminopropylmethacrylamide (APMAAm) for over 20 passages in chemically-defined mTeSR™1 media and demonstrated pluripotency of multiple hESC lines with immunostaining and quantitative RT-PCR studies. Results for hESC proliferation and pluripotency markers were both qualitatively and quantitatively similar to cells cultured on Matrigel™-coated substrates. Mechanistically, it was resolved that bovine serum albumin (BSA) in the mTeSR™1 media was critical for cell adhesion on APMAAm hydrogel interfaces. This study uniquely identified a robust long-term culture surface for the self-renewal of hESCs without the use of biologic coatings (e.g., peptides, proteins, or Matrigel™) in completely chemically-defined media that employed practical culturing techniques amenable to clinical-scale cell expansion.
authors.affiliation: Department of Bioengineering, University of California at Berkeley, Berkeley, CA 94720, United States.
authors.forename: Elizabeth F
authors.initials: EF
authors.lastname: Irwin
authors.forename: Rohini
authors.initials: R
authors.lastname: Gupta
authors.forename: Derek C
authors.initials: DC
authors.lastname: Dashti
authors.forename: Kevin E
authors.initials: KE
authors.lastname: Healy
mesh_terms: D000178:Acrylamides / Q000737:chemistry; D000327:Adsorption; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology*; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D008422:Materials Testing; D011108:Polymers / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013499:Surface Properties
chemical_list: D000178:Acrylamides; D001672:Biocompatible Materials; D003470:Culture Media; D020100:Hydrogels; D011108:Polymers; D012710:Serum Albumin, Bovine; C045985:methacrylamide