title: Fetal human hematopoietic stem cells can differentiate sequentially into neural stem cells and then astrocytes in vitro.
abstract: In some rodent models, there is evidence that hematopoietic stem cells (HSC) can differentiate into neural cells. However, it is not known whether humans share this potential, and, if so, what conditions are sufficient for this transdifferentiation to occur. We addressed this question by assessing the ability of fetal human liver CD34(+)/CD133(+)/CD3(-) hematopoietic stem cells to generate neural cells and astrocytes in culture. We cultured fetal liver-derived hematopoietic stem cells in human astrocyte culture-conditioned medium or using a method wherein growing human astrocytes were separated from cultured, nonadherent hematopoietic stem cells by a semipermeable membrane in a double-chamber co-culture system. Hematopoietic stem cell cultures were probed for neural progenitor cell marker expression (nestin and bone morphogenic protein-2 [BMP-2]) during growth in both culture conditions. RT-PCR, western blotting, and immunocytochemistry assays showed that cells cultured in either condition expressed nestin mRNA and protein and BMP-2 mRNA. HSC similarly cultured in nonconditioned medium or in the absence of astrocytes did not express either marker. Cells expressing these neural markers were transferred and cultured on poly-D-lysine-coated dishes with nonconditioned growth medium for further study. Immunocytochemistry demonstrated that these cells differentiated into astrocytes after 8 days in culture as indicated by their morphology and expression of the astrocytic markers glial fibrillary acidic protein (GFAP) and S100, as well as by their rate of proliferation, which was identical to that of freshly isolated fetal brain astrocytes. These findings demonstrate that neural precursor gene expression can be induced when human hematopoietic stem cells are exposed to a suitable microenvironment. Furthermore, the neural stem cells generated in this environment can then differentiate into astrocytes. Therefore, human hematopoietic stem cells may be an alternative resource for generation of neural stem cells for therapy of central nervous system defects resulting from disease or trauma.
authors.affiliation: Department of Pediatrics, Wayne State University School of Medicine and Children's Research Center of Michigan, Detroit, MI 48201, USA. HsiaoNan.Hao@wayne.edu
authors.forename: Hsiao-Nan
authors.initials: HN
authors.lastname: Hao
authors.forename: Jane
authors.initials: J
authors.lastname: Zhao
authors.forename: Ronald L
authors.initials: RL
authors.lastname: Thomas
authors.forename: Graham C
authors.initials: GC
authors.lastname: Parker
authors.forename: William D
authors.initials: WD
authors.lastname: Lyman
mesh_terms: D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34 / Q000096:biosynthesis; D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism; D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000378:metabolism; D017252:CD3 Complex / Q000096:biosynthesis; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology; D005434:Flow Cytometry; D006023:Glycoproteins / Q000096:biosynthesis; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins; D064231:Nestin; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D010455:Peptides; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors; D016212:Transforming Growth Factor beta
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D017252:CD3 Complex; D017077:Culture Media, Conditioned; D006023:Glycoproteins; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; C000606023:PROM1 protein, human; D010455:Peptides; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta