title: Collagen IV significantly enhances migration and transplantation of embryonic stem cells: involvement of α2β1 integrin-mediated actin remodeling.
abstract: Embryonic stem (ES) cell transplantation represents a potential means for the treatment of degenerative diseases and injuries. As appropriate distribution of transplanted ES cells in the host tissue is critical for successful transplantation, the exploration of efficient strategies to enhance ES cell migration is warranted. In this study we investigated ES cell migration under the influence of various extracellular matrix (ECM) proteins, which have been shown to stimulate cell migration in various cell models with unclear effects on ES cells. Using two mouse ES (mES) cell lines, ESC 26GJ9012-8-2 and ES-D3 GL, to generate embryoid bodies (EBs), we examined the migration of differentiating cells from EBs that were delivered onto culture surfaces coated with or without collagen I, collagen IV, Matrigel, fibronectin, and laminin. Among these ECM proteins, collagen IV exhibited maximal migration enhancing effect. mES cells expressed α2 and β1 integrin subunits and the migration enhancing effect of collagen IV was prevented by RGD peptides as well as antibodies against α2 and β1 integrins, indicating that the enhancing effect of collagen IV on cell migration was mediated by α2β1 integrin. Furthermore, staining of actin cytoskeleton that links to integrins revealed well-developed stress fibers and long filopodia in mES cells cultured on collagen IV, and the actin-disrupting cytochalasin D abolished the collagen IV-enhanced cell migration. In addition, pretreatment of undifferentiated or differentiated mES cells with collagen IV resulted in improved engraftment and growth after transplantation into the subcutaneous tissue of nude mice. Finally, collagen IV pretreatment of osteogenically differentiated mES cells increased osteogenic differentiation-like tissue and decreased undifferentiation-like tissue in the grafts grown after transplantation. Our results demonstrated that collagen IV significantly enhanced the migration of differentiating ES cells through α2β1 integrin-mediated actin remodeling and could promote ES cell transplantation efficiency, which may be imperative to stem cell therapy.
authors.affiliation: Department of Obstetrics and Gynecology, Taipei Veterans General Hospital, Taipei, Taiwan.
authors.forename: Hsin-Yang
authors.initials: HY
authors.lastname: Li
authors.forename: Chen-Yi
authors.initials: CY
authors.lastname: Liao
authors.forename: Kun-Hsiung
authors.initials: KH
authors.lastname: Lee
authors.forename: Hung-Chi
authors.initials: HC
authors.lastname: Chang
authors.forename: Yi-Jen
authors.initials: YJ
authors.lastname: Chen
authors.forename: Kuan-Chong
authors.initials: KC
authors.lastname: Chao
authors.forename: Sheng-Ping
authors.initials: SP
authors.lastname: Chang
authors.forename: Hsin-Yi
authors.initials: HY
authors.lastname: Cheng
authors.forename: Chia-Ming
authors.initials: CM
authors.lastname: Chang
authors.forename: Yuh-Lih
authors.initials: YL
authors.lastname: Chang
authors.forename: Shih-Chieh
authors.initials: SC
authors.lastname: Hung
authors.forename: Yen-Jen
authors.initials: YJ
authors.lastname: Sung
authors.forename: Shih-Hwa
authors.initials: SH
authors.lastname: Chiou
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects; D000199:Actins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D000906:Antibodies / Q000276:immunology; D002454:Cell Differentiation; D002465:Cell Movement; D024141:Collagen Type IV / Q000494:pharmacology*; D015638:Cytochalasin D / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000637:transplantation*; D016326:Extracellular Matrix Proteins / Q000494:pharmacology; D038982:Integrin alpha2beta1 / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D009842:Oligopeptides / Q000494:pharmacology
chemical_list: D000199:Actins; D000906:Antibodies; D024141:Collagen Type IV; D016326:Extracellular Matrix Proteins; D038982:Integrin alpha2beta1; D009842:Oligopeptides; D015638:Cytochalasin D; C047981:arginyl-glycyl-aspartic acid