title: The Rho kinase inhibitor Y-27632 facilitates the differentiation of bone marrow mesenchymal stem cells.
abstract: The selective in vitro expansion and differentiation of multipotent stem cells are critical steps in cell-based regenerative therapies, but technical challenges have limited cell yield and thus the success of these potential treatments. The Rho GTPases and downstream Rho kinases (Rho coiled-coil kinases or ROCKs) are central regulators of cytoskeletal dynamics during the cell cycle and thus help determine the balance between stem cells self-renewal, lineage commitment, and apoptosis. Here, we examined if suppression of ROCK signaling enhances the efficacy of bone marrow-derived mesenchymal stem cells (BMSCs) differentiation into neurons and neuroglial cells. BMSCs were cultured in epidermal growth factor (EGF, 10 µg/l) and basic fibroblastic growth factor (bFGF, 10 µg/l) in the presence or absence of the Rho kinase inhibitor Y-27632 (10 µM). The expression levels of neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) were detected by immunofluorescence and Western blotting. The average number of NSE-positive cells increased from 83.20 ± 8.677 (positive ratio 0.2140 ± 0.0119) to 109.20 ± 8.430 (positive ratio 0.3193 ± 0.0161) per visual field in the presence of Y-27632, while GFAP-positive cell number increased from 96.30 ± 8.486 (positive ratio 0.18 ± 0.0152) to 107.50 ± 8.683 (positive ratio 0.27 ± 0.0115) (P < 0.05 for both). Both NSE and GFAP protein expression levels were enhanced significantly by Y-27632 treatment (NSE: 0.74 ± 0.05 vs. 1.03 ± 0.06; GFAP: 0.64 ± 0.08 vs. 0.97 ± 0.05, both P < 0.01) as indicated by Western blots. The Rho kinase inhibitor Y-27632 concomitant with EGF and bFGF stimulation promotes BMSC differentiation into neural cells. Control of Rho kinase activity may enhance the efficiency of stem cell-based treatments for neurodegenerative diseases.
authors.affiliation: Emergency Center, The Affiliated Hospital of Xuzhou Medical College, First Aid and Relief Medical Department of Xuzhou Medical College, Xuzhou, 221002, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Liu
authors.forename: Zhengzheng
authors.initials: Z
authors.lastname: Zhang
authors.forename: Xianliang
authors.initials: X
authors.lastname: Yan
authors.forename: He
authors.initials: H
authors.lastname: Liu
authors.forename: Licai
authors.initials: L
authors.lastname: Zhang
authors.forename: Aiming
authors.initials: A
authors.lastname: Yao
authors.forename: Chengcheng
authors.initials: C
authors.lastname: Guo
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Liu
authors.forename: Tie
authors.initials: T
authors.lastname: Xu
mesh_terms: D000577:Amides / Q000494:pharmacology*; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D008297:Male; D059630:Mesenchymal Stem Cells / Q000201:enzymology / Q000502:physiology*; D009457:Neuroglia / Q000502:physiology; D009474:Neurons / Q000502:physiology; D011725:Pyridines / Q000494:pharmacology*; D017207:Rats, Sprague-Dawley; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D000577:Amides; D011725:Pyridines; C108830:Y 27632; D054460:rho-Associated Kinases