EGFP expression controlled by GFAP promoter in mesenchymal cells: an efficient tool for glial lineage selection and transplantation.
In order to demonstrate a new method to label and select enough glial cells from induced MSCs to provide cells for cell therapy, MSCs were induced with Beta-mercaptoethanol followed by retinoic acid, forskolin, basic-FGF, PDGF and heregulin. Induced MSCs were transfected with reconstructed vector pGFAP-EGFP by inserting GFAP promotor into pEGFP-N3 to substitute CMV promotor. Living cells against G418 were enriched and checked by flowcytometry. EGFP expressing cells were sorted and used for transplantation in vivo. Immunoelectronmicroscopy was accomplished using anti-EGFP to relocalize the transplanted cells. Almost all MSCs took on phenotypes of glial cells after induction, expressing S100 and GFAP. The EGFP expression rate of survived MSCs against G418 was 82.74%. Glial cells expressing EGFP accumulated mainly around the damaged nerve fibers. MSCs were relocalized by immunoelectronmicroscopy and remyelination was observed. EGFP expression controlled by GFAP promoter in mesenchymal cells was an efficient tool for glial lineage selection and transplantation. Induced MSCs can promote nerve regeneration by participating remyelination.
Zhang
Pei-Xun
PX
People's Hospital, Peking University, Beijing, People's Republic of China.
He
Xiang-Jun
XJ
Zhao
Fu-Qiang
FQ
Du
Chan
C
Fu
Zhong-Guo
ZG
Zhang
Dian-Ying
DY
Zhang
Hong-Bo
HB
Jiang
Bao-Guo
BG
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D005904:Glial Fibrillary Acidic Protein; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D009457:Neuroglia / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012584:Sciatic Nerve / Q000293:injuries; D020196:Trauma, Nervous System / Q000628:therapy*
D005904:Glial Fibrillary Acidic Protein; D049452:Green Fluorescent Proteins