title: Neural progenitor cell survival in mouse brain can be improved by co-transplantation of helper cells expressing bFGF under doxycycline control.
abstract: Cell-based therapy of neurological disorders is hampered by poor survival of grafted neural progenitor cells (NPCs). We hypothesized that it is possible to enhance the survival of human NPCs (ReNcells) by co-transplantation of helper cells expressing basic fibroblast growth factor (bFGF) under control of doxycycline (Dox). 293 cells or C17.2 cells were transduced with a lentiviral vector encoding the fluorescent reporter mCherry and bFGF under tetracycline-regulated transgene expression (Tet-ON). The bFGF secretion level in the engineered helper cells was positively correlated with the dose of Dox (Pearson correlation test; r=0.95 and 0.99 for 293 and C17.2 cells, respectively). Using bioluminescence imaging (BLI) as readout for firefly luciferase-transduced NPC survival, the addition of both 293-bFGF and C17.2-bFGF helper cells was found to significantly improve cell survival up to 6-fold in vitro, while wild-type (WT, non-transduced) helper cells had no effect. Following co-transplantation of 293-bFGF or C17.2-bFGF cells in the striatum of Rag2(-/-) immunodeficient mice, in vivo human NPC survival could be significantly improved as compared to no helper cells or co-transplantation of WT cells for the first two days after co-transplantation. This enhancement of survival in C17.2-bFGF group was not achieved without Dox administration, indicating that the neuroprotective effect was specific for bFGF. The present results warrant further studies on the use of engineered helper cells, including those expressing other growth factors injected as mixed cell populations.
authors.affiliation: Russell H. Morgan Dept. of Radiology and Radiological Science, Division of MR Research, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Yajie
authors.initials: Y
authors.lastname: Liang
authors.forename: Louise
authors.initials: L
authors.lastname: Ågren
authors.forename: Agatha
authors.initials: A
authors.lastname: Lyczek
authors.forename: Piotr
authors.initials: P
authors.lastname: Walczak
authors.forename: Jeff W M
authors.initials: JW
authors.lastname: Bulte
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology* / Q000601:surgery*; D002450:Cell Communication; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D017690:Cell Transplantation; D002478:Cells, Cultured; D018920:Coculture Techniques; D004268:DNA-Binding Proteins / Q000172:deficiency; D004305:Dose-Response Relationship, Drug; D004318:Doxycycline / Q000494:pharmacology*; D005346:Fibroblast Growth Factors / Q000096:biosynthesis / Q000378:metabolism*; D049454:Firefly Luciferin / Q000008:administration & dosage / Q000378:metabolism; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008163:Luminescent Measurements; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D058953:Neural Stem Cells / Q000187:drug effects* / Q000502:physiology*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D004268:DNA-Binding Proteins; D004318:Doxycycline; D005346:Fibroblast Growth Factors; D049454:Firefly Luciferin; D008164:Luminescent Proteins; D001616:beta-Galactosidase; D000097573:Red Fluorescent Protein; C499308:Rag2 protein, mouse
keywords: Bioluminescent imaging; Cell survival; Neural progenitor cells; Transplantation; bFGF