Document Detail


Isolation, characterization and preclinical development of human glial-restricted progenitor cells for treatment of neurological disorders.
MedLine Citation:
PMID:  20455649     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
AIM: Glial-restricted progenitor cells (GRPs), a neural cell population that gives rise to astrocytes and oligodendrocytes both in vitro and in vivo, hold great promise as a cellular therapeutic for the treatment of demyelinating and neurodegenerative diseases of the CNS. The manufacturing and characterization protocols of human-derived GRPs (hGRPs; trade name Q-Cells) for use in a clinical setting that adhere to rigorous standards for their isolation, propagation, characterization and storage are presented. MATERIALS & METHODS: hGRPs, defined by their immunoreactivity with A2B5 antibodies, were isolated from fetal cadaver forebrain tissue of mice 17-24 weeks gestational age using Miltenyi paramagnetic bead cell separation technology. GRPs were grown in a defined xenobiotic-free medium for 6 days. At harvest, hGRPs were characterized using immunocytochemical techniques. Long-term cryopreservation and storage conditions, and viability upon freeze-thaw were determined. The phenotypic differentiation potential of hGRPs was determined by implantation experiments into the CNS of shiverer mice. RESULTS: hGRPs were isolated from over 50 neural tissues of either sex during gestational ages of 17-24 weeks. Cells expanded out to 6 days in vitro in a xenobiotic-free medium demonstrated very consistent immunocytochemical profiles. No residual antibody used in the purification process was detected after 6 days of growth in vitro. GRPs could be frozen at up to 24 million cells/ml and were over 70% viable upon freeze-thaw. Thawed hGRPs transplanted into the brain of the dysmyelinated shiverer mouse model were observed to differentiate into both glial fibrillary acidic protein-positive astrocytes and myelin basic protein-positive oligodendrocytes; no human-derived NeuN-positive neuronal cells were observed and no abnormal cell proliferation was observed. CONCLUSION: We demonstrate that hGRPs can be consistently obtained, propagated, cryopreserved and characterized using protocols that can be transferred to a good laboratory practice/good manufacturing practice setting for the manufacture of clinical-grade hGRP cellular therapeutics. Functional data demonstrate that cells manufactured under these conditions are able to differentiate into appropriate cellular phenotypes in an animal model of dysmyelination.
Authors:
Robert W Sandrock; Will Wheatley; Cynthia Levinthal; Jennifer Lawson; Brooke Hashimoto; Mahendra Rao; James T Campanelli
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Regenerative medicine     Volume:  5     ISSN:  1746-076X     ISO Abbreviation:  Regen Med     Publication Date:  2010 May 
Date Detail:
Created Date:  2010-05-11     Completed Date:  2010-10-12     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101278116     Medline TA:  Regen Med     Country:  England    
Other Details:
Languages:  eng     Pagination:  381-94     Citation Subset:  IM    
Affiliation:
Q Therapeutics, Inc., 615 Arapeen Drive, Suite 102, Salt Lake City, UT 84108, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Differentiation
Cell Transplantation / methods*
Epitopes / chemistry
Female
Flow Cytometry / methods
Humans
Immunohistochemistry / methods
Male
Mice
Neurodegenerative Diseases / metabolism*,  therapy
Neuroglia / cytology*
Prosencephalon / metabolism
Regenerative Medicine / methods
Stem Cells / cytology*
Chemical
Reg. No./Substance:
0/Epitopes

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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