In Vitro Labeling Strategies for Identifying Primary Neural Tissue and a Neuronal Cell Line after Transplantation in the Cns
Autor: | Stephen M. Onifer, Holets Vr, Whittemore, White La |
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Jazyk: | angličtina |
Rok vydání: | 1993 |
Předmět: |
Male
0301 basic medicine Serotonin Cell division 5 7-Dihydroxytryptamine Biomedical Engineering lcsh:Medicine Biology Transfection Cell Line Rats Sprague-Dawley 03 medical and health sciences 0302 clinical medicine Fetal Tissue Transplantation Pregnancy In vivo Escherichia coli Animals Brain Tissue Transplantation Phytohemagglutinins Cells Cultured Fluorescent Dyes Neurons Transplantation Raphe Graft Survival lcsh:R Cell Biology beta-Galactosidase Embryonic stem cell Molecular biology In vitro Rats 030104 developmental biology Genes Bacterial Rats Inbred Lew Cell culture Raphe Nuclei Female Cell Division 030217 neurology & neurosurgery |
Zdroj: | Cell Transplantation, Vol 2 (1993) |
ISSN: | 1555-3892 0963-6897 |
Popis: | Potential labels for identifying embryonic raphe neurons and a clonal, neuronally differentiating, raphe-derived cell line, RN33B, in CNS transplantation studies were tested by first characterizing the labels in vitro. The labels that were tested included 4,6-diamidino-2-phenylindole hydrochloride, 1,1′-dioctadecyl-3,3,3′-tetramethylindocarbocyanine perchlorate, the Escherichia coli lacZ gene, Fast Blue, Fluoro-Gold, fluorescein-conjugated latex microspheres, fluorescein isothiocyanate-conjugated or nonconjugated Phaseolus vulgaris leucoagglutinin, methyl o-(6-amino-3′-imino-3H-xanthen-9-yl) benzoate monohydrochloride, or tetanus toxin C fragment. Subsequently, the optimal in vitro labels for embryonic raphe neurons and for RN33B cells were characterized in vivo after CNS transplantation. In vitro, 1,1-dioctadecyl-3,3,3′-tetramethylindocarbocyanine perchlorate (DiI) optimally labeled embryonic neurons. The Escherichia coli lacZ gene optimally labeled RN33B cells. Most labels were rapidly diluted in cultures of embryonic astrocytes and proliferating RN33B cells. Some labels were toxic and were often retained in cellular debris. In vivo, DiI was visualized in transplanted, DiI-labeled raphe neurons, but not in astrocytes up to 1 mo posttransplant. DiI-labeled host cells were seen after transplantation of lysed, DiI-labeled cells. β-Galactosidase was visualized in transplanted, Escherichia coli lacZ gene-labeled RN33B cells after 15 days in vivo. No β-galactosidase was seen in host cells after transplantation of lysed, lacZ-labeled RN33B cells. The results demonstrate that labels for use in CNS transplantation studies should be optimized for the specific population of donor cells under study, with the initial step being characterization in vitro followed by in vivo analysis. Appropriate controls for false-positive labeling of host cells should always be assessed. |
Databáze: | OpenAIRE |
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