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Purification of human fetal hippocampal neurons by flow cytometry for transplantation.
Chiu, F C; Potter, P E; Rozental, R.
Afiliação
  • Chiu FC; Department of Anesthesiology, Montefiore Medical Center and Albert Einstein College of Medicine, Bronx, New York 10467, USA.
Methods ; 16(3): 260-7, 1998 Nov.
Article em En | MEDLINE | ID: mdl-10071065
ABSTRACT
We have established primary cultures, highly enriched in neurons, from the hippocampus of human fetal brains at 20-23 gestational weeks. More than 80% of cells were viable when seeded. Neurons were isolated from primary cultures by flow cytometry to a high degree of purity, as demonstrated by immunocytochemical staining. FACS scanning analysis using a DNA-staining dye showed that hippocampal neurons did not divide in culture. To demonstrate that FACS-sorted neurons can be transplanted and integrated into the host brain, neuron-enriched primary culture from human fetal striatum was infected with a viral-mediated vector containing a reporter gene, beta-galactosidase. Striatal neurons were subsequently purified by flow cytometry and transplanted into the striatum of rats. Following transplantation, the rat brains were processed for beta-galactosidase histochemistry and electron microscopy. Beta-galactosidase expression indicates that transplanted human neurons survived in the host and were metabolically active. The transplanted neurons received synaptic inputs, as judged from the presence of presynaptic terminals on their surface. Our study demonstrates connectivity between transplanted human fetal primary neurons and host tissue at the ultrastructural level. Our results support the feasibility of ultimately transplanting neurons into humans as a possible treatment for recovery of the nervous system (e.g., neurodegenerative diseases).
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Transplante de Tecido Encefálico / Citometria de Fluxo / Hipocampo / Neurônios Limite: Animals / Humans Idioma: En Revista: Methods Assunto da revista: BIOQUIMICA Ano de publicação: 1998 Tipo de documento: Article País de afiliação: Estados Unidos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Transplante de Tecido Encefálico / Citometria de Fluxo / Hipocampo / Neurônios Limite: Animals / Humans Idioma: En Revista: Methods Assunto da revista: BIOQUIMICA Ano de publicação: 1998 Tipo de documento: Article País de afiliação: Estados Unidos