title: Improving viability and transfection efficiency with human umbilical cord wharton's jelly cells through use of a ROCK inhibitor.
abstract: Differentiating stem cells using gene delivery is a key strategy in tissue engineering and regenerative medicine applications. Nonviral gene delivery bypasses several safety concerns associated with viral gene delivery; however, leading nonviral techniques, such as electroporation, subject cells to high stress and can result in poor cell viabilities. Inhibition of Rho-associated coiled-coil kinase (ROCK) has been shown to mitigate apoptotic mechanisms associated with detachment and freezing of induced pluripotent stem cells and embryonic stem cells; however, inhibiting ROCK in mesenchymal stromal cells (MSCs) for improving gene delivery applications has not been reported previously. In this study, we hypothesized that ROCK Inhibitor (RI) would improve cell viability and gene expression in primary human umbilical cord mesenchymal stromal cells (hUCMSCs) when transfected via Nucleofection™. As hypothesized, the pre-treatment and post-treatment of hUCMSCs transfected via nucleofection with Y-27632-RI significantly improved survival rates of hUCMSCs and gene expression as measured by green fluorescent protein intensity. This study provides the first comparative look at the effect of Y-27632-RI on hUCMSCs that underwent transfection via nucleofection and shows that using Y-27632-RI in concert with nucleofection could greatly enhance the utility of differentiating and reprogramming hUCMSCs for tissue engineering applications.
authors.affiliation: 1 Bioengineering Program, University of Kansas , Lawrence, KS, 66045.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Mellott
authors.forename: Megan E
authors.initials: ME
authors.lastname: Godsey
authors.forename: Heather E
authors.initials: HE
authors.lastname: Shinogle
authors.forename: David S
authors.initials: DS
authors.lastname: Moore
authors.forename: M Laird
authors.initials: ML
authors.lastname: Forrest
authors.forename: Michael S
authors.initials: MS
authors.lastname: Detamore
mesh_terms: D000577:Amides / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002470:Cell Survival; D005260:Female; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology*; D014162:Transfection; D014470:Umbilical Cord / Q000166:cytology / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
chemical_list: D000577:Amides; D047428:Protein Kinase Inhibitors; D011725:Pyridines; C108830:Y 27632; D054460:rho-Associated Kinases