title: Transplantation of mouse embryonic stem cell-derived oligodendrocytes in the murine model of globoid cell leukodystrophy.
abstract: BACKGROUND
  Globoid cell leukodystrophy (GLD) is a severe disorder of the central and peripheral nervous system caused by the absence of galactocerebrosidase (GALC) activity. Cell-based therapies are highly promising strategies for GLD. In this study, G-Olig2 mouse embryonic stem cells (ESCs) were induced into oligodendrocyte progenitor cells (OPCs) and were implanted into the brains of twitcher mice, an animal model of GLD, to explore the therapeutic potential of the cells.
  
  
  METHODS
  The G-Olig2 ESCs were induced into OPCs by using cytokines and a multi-step differentiation procedure. Oligodendrocyte markers were detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. The toxicity of psychosine to OPCs was determined by a cell proliferation assay kit. The GALC level of OPCs was also examined. OPCs were labeled with Dir and transplanted into the brains of twitcher mice. The transplanted cells were detected by in-Vivo Multispectral Imaging System and real-time PCR. The physiological effects of twitcher mice were assessed.
  
  
  RESULTS
  Oligodendrocyte markers were expressed in OPCs, and 76%±5.76% of the OPCs were enhanced green fluorescent protein (eGFP)-positive, eGFP was driven by the Olig2 promoter. The effect of psychosine on cell viability indicated that OPCs were more resistant to psychosine toxicity. The GALC level of OPCs was 10.0±1.23 nmol/hour per mg protein, which was significantly higher than other cells. Dir-labeled OPCs were injected into the forebrain of post-natal day 10 twitcher mice. The transplanted OPCs were myelin basic protein (MBP)-positive and remained along the injection tract as observed by fluorescent microscopy. The level of the Dir fluorescent signal and eGFP mRNA significantly decreased at days 10 and 20 after injection, as indicated by in-Vivo Multispectral Imaging System and real-time PCR. Because of poor cell survival and limited migration ability, there was no significant improvement in brain GALC activity, MBP level, life span, body weight, and behavioral deficits of twitcher mice.
  
  
  CONCLUSIONS
  ESC-derived OPC transplantation was not sufficient to reverse the clinical course of GLD in twitcher mice.
authors.affiliation: Department of Gastroenterology, Nantong University Affiliated Hospital, 20 Xi Si Road, Nantong, Jiangsu, 226001, China. kxl720724@yahoo.com.
authors.forename: Xiao Ling
authors.initials: XL
authors.lastname: Kuai
authors.affiliation: Department of Gastroenterology, Nantong University Affiliated Hospital, 20 Xi Si Road, Nantong, Jiangsu, 226001, China. nirz@163.com.
authors.forename: Run Zhou
authors.initials: RZ
authors.lastname: Ni
authors.affiliation: Department of Gastroenterology, Nantong University Affiliated Hospital, 20 Xi Si Road, Nantong, Jiangsu, 226001, China. zhou.guoxiong@aliyun.com.
authors.forename: Guo Xiong
authors.initials: GX
authors.lastname: Zhou
authors.affiliation: Department of Gastroenterology, Nantong University Affiliated Hospital, 20 Xi Si Road, Nantong, Jiangsu, 226001, China. mzb63@163.com.
authors.forename: Zheng Biao
authors.initials: ZB
authors.lastname: Mao
authors.affiliation: Department of Gastroenterology, Nantong University Affiliated Hospital, 20 Xi Si Road, Nantong, Jiangsu, 226001, China. Zhangjf05090@163.com.
authors.forename: Jian Feng
authors.initials: JF
authors.lastname: Zhang
authors.affiliation: Department of Gastroenterology, Nantong University Affiliated Hospital, 20 Xi Si Road, Nantong, Jiangsu, 226001, China. jsntyi@163.com.
authors.forename: Nan
authors.initials: N
authors.lastname: Yi
authors.affiliation: Department of Gastroenterology, Nantong University Affiliated Hospital, 20 Xi Si Road, Nantong, Jiangsu, 226001, China. joezyz@hotmail.com.
authors.forename: Zhao Xiu
authors.initials: ZX
authors.lastname: Liu
authors.affiliation: Department of Gastroenterology, Nantong University Affiliated Hospital, 20 Xi Si Road, Nantong, Jiangsu, 226001, China. shaonanma@163.com.
authors.forename: Nan
authors.initials: N
authors.lastname: Shao
authors.affiliation: Department of Gastroenterology, Nantong University Affiliated Hospital, 20 Xi Si Road, Nantong, Jiangsu, 226001, China. niwenkai@sina.com.
authors.forename: Wen Kai
authors.initials: WK
authors.lastname: Ni
authors.affiliation: Department of General Surgery, Nantong University Affiliated Hospital, 20 Xi Si Road, Nantong, Jiangsu, 226001, China. alexxs@139.com.
authors.forename: Zhi Wei
authors.initials: ZW
authors.lastname: Wang
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001921:Brain / Q000473:pathology / Q000601:surgery; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D002465:Cell Movement; D002470:Cell Survival; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D004195:Disease Models, Animal; D005698:Galactosylceramidase / Q000378:metabolism; D007965:Leukodystrophy, Globoid Cell / Q000628:therapy*; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000637:transplantation*; D009186:Myelin Sheath / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology / Q000637:transplantation*; D011609:Psychosine / Q000378:metabolism; D033581:Stem Cell Transplantation; D017211:Treatment Failure
chemical_list: D015415:Biomarkers; D011609:Psychosine; D005698:Galactosylceramidase