title: Markers of pluripotency and differentiation in human neural precursor cells derived from embryonic stem cells and CNS tissue.
abstract: Cell transplantation therapies for central nervous system (CNS) deficits such as spinal cord injury (SCI) have been shown to be effective in several animal models. One cell type that has been transplanted is neural precursor cells (NPCs), for which there are several possible sources. We have studied NPCs derived from human embryonic stem cells (hESCs) and human fetal CNS tissue (hfNPCs), cultured as neurospheres, and the expression of pluripotency and neural genes during neural induction and in vitro differentiation. mRNA for the pluripotency markers Nanog, Oct-4, Gdf3, and DNMT3b were downregulated during neural differentiation of hESCs. mRNA for these markers was found in nonpluripotent hfNPC at higher levels compared to hESC-NPCs. However, Oct-4 protein was found in hESC-NPCs after 8 weeks of culture, but not in hfNPCs. Similarly, SSEA-4 and CD326 were only found in hESC-NPCs. NPCs from both sources differentiated as expected to cells with typical features of neurons and astrocytes. The expressions of neuronal markers in hESC-NPCs were affected by the composition of cell culture medium, while this did not affect hfNPCs. Transplantation of hESC-NPC or hfNPC neurospheres into immunodeficient mouse testis or subcutaneous tissue did not result in tumor formation. In contrast, typical teratomas appeared in all animals after transplantation of hESC-NPCs to injured or noninjured spinal cords of immunodeficient rats. Our data show that transplantation to the subcutaneous tissue or the testes of immunodeficient mice is not a reliable method for evaluation of the tumor risk of remaining pluripotent cells in grafts.
authors.affiliation: Regea-Institute for Regenerative Medicine, University of Tampere and Tampere University Hospital, Tampere, Finland.
authors.forename: M
authors.initials: M
authors.lastname: Sundberg
authors.forename: P-H
authors.initials: PH
authors.lastname: Andersson
authors.forename: E
authors.initials: E
authors.lastname: Åkesson
authors.forename: J
authors.initials: J
authors.lastname: Odeberg
authors.forename: L
authors.initials: L
authors.lastname: Holmberg
authors.forename: J
authors.initials: J
authors.lastname: Inzunza
authors.forename: S
authors.initials: S
authors.lastname: Falci
authors.forename: J
authors.initials: J
authors.lastname: Öhman
authors.forename: R
authors.initials: R
authors.lastname: Suuronen
authors.forename: H
authors.initials: H
authors.lastname: Skottman
authors.forename: K
authors.initials: K
authors.lastname: Lehtimäki
authors.forename: O
authors.initials: O
authors.lastname: Hovatta
authors.forename: S
authors.initials: S
authors.lastname: Narkilahti
authors.forename: E
authors.initials: E
authors.lastname: Sundström
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D002490:Central Nervous System / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D005333:Fetus / Q000166:cytology; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D016513:Mice, SCID; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013119:Spinal Cord Injuries / Q000473:pathology / Q000628:therapy; D033581:Stem Cell Transplantation; D040521:Subcutaneous Tissue; D013737:Testis / Q000166:cytology
chemical_list: D015415:Biomarkers; D012333:RNA, Messenger