title: A method to generate enhanced GFP+ chimeric mice to study the role of bone marrow-derived cells in the eye.
abstract: GFP-chimeric mice are important tools to study the role of bone marrow-derived cells in eye physiology. A method is described to generate GFP-chimeric mice using whole-body, sub-lethal radiation (600 rad) of wild-type C57BL/6 recipients followed by tail vein injection of bone marrow cells derived from GFP+ (GFP-transgenic C57/BL/6-Tg(UBC-GFP)30 Scha/J) mice. This method yields stable GFP+ chimeras with greater than 95% chimerism (range 95-99%), achieved within one month of bone marrow transfer confirmed by microscopy and fluorescence-assisted cell sorting (FACS) analysis, with lower mortality after irradiation than prior methods. To demonstrate the efficacy of GFP+ bone marrow chimeric mice, the role of circulating GFP+ bone marrow-derived cells in myofibroblast generation after irregular photo-therapeutic keratectomy (PTK) was analyzed. Many SMA+ myofibroblasts that were generated at one month after PTK were derived from GFP+ bone marrow-derived cells. The GFP+ bone marrow chimeric mouse provides an excellent model for studying the role of bone marrow-derived cells in corneal wound healing, glaucoma surgery, optic nerve head pathology and retinal pathophysiology and wound healing.
authors.affiliation: Cole Eye Institute, Cleveland Clinic, 9500 Euclid Ave, Cleveland, OH 44195, USA.
authors.forename: Vivek
authors.initials: V
authors.lastname: Singh
authors.forename: Ritika
authors.initials: R
authors.lastname: Jaini
authors.forename: André A M
authors.initials: AA
authors.lastname: Torricelli
authors.forename: Vincent K
authors.initials: VK
authors.lastname: Tuohy
authors.forename: Steven E
authors.initials: SE
authors.lastname: Wilson
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D016026:Bone Marrow Transplantation / Q000379:methods*; D003315:Cornea / Q000473:pathology / Q000601:surgery; D065306:Corneal Injuries; D004195:Disease Models, Animal; D005131:Eye Injuries / Q000378:metabolism / Q000473:pathology / Q000601:surgery*; D005260:Female; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D014945:Wound Healing
keywords: bone marrow-derived cells; chimeric; fibrosis; green fluorescent protein; mice; myofibroblasts; wound healing