Human fetal myelinated organotypic cultures.
We have previously reported the establishment of organotypic cultures derived from human fetal brain tissue. Although these cultures permit the testing of multiple hypotheses about normal human neurodevelopment and neuropathologic conditions, they have the limitation of not being myelinated and therefore preclude the study of questions related to myelinogenesis and diseases of myelin. In the current communication, we describe recent developments that allow us to overcome this limitation and permit the establishment of a myelinated organotypic culture model. Sections of dorsal column dissected from the lumbar spinal cord of human fetuses ranging in age 21-23 weeks of gestation were placed in culture. The explants were maintained for up to 12 weeks during which time they were characterized and shown to express a number of CNS cell-type-specific markers including glial fibrillary acidic protein (astrocytes), nerve growth factor receptor and neurofilament protein (neurons), CD68 (microglia), and myelin basic protein, HNK-1 and galactocerebroside (oligodendrocytes). In addition, lectin histochemistry using Ricinus communis agglutinin-1 detected microglia and endothelial cells. Upon explantation, abundant myelin was seen by electron microscopy in the cultures. Although during the culture period there was degradation of myelin, there was also evidence of maintenance of intact myelin sheaths around small caliber axons and de novo myelin synthesis. This model system may permit the further use of human organotypic cultures to investigate issues related to neurodevelopment and to pathologic conditions including those relevant to dysmyelination and demyelination.
Lyman
W D
WD
Department of Pathology, Albert Einstein College of Medicine, Bronx, NY 10461.
Hatch
W C
WC
Pousada
E
E
Stephney
G
G
Rashbaum
W K
WK
Weidenheim
K M
KM
D001253:Astrocytes / Q000166:cytology* / Q000648:ultrastructure; D001369:Axons / Q000648:ultrastructure; D015415:Biomarkers; D001921:Brain / Q000166:cytology* / Q000196:embryology / Q000648:ultrastructure; D005260:Female; D005333:Fetus; D005865:Gestational Age; D006801:Humans; D008648:Mesoderm / Q000166:cytology / Q000648:ultrastructure; D008854:Microscopy, Electron; D009186:Myelin Sheath / Q000648:ultrastructure*; D009413:Nerve Fibers, Myelinated / Q000648:ultrastructure*; D016501:Neurites / Q000648:ultrastructure; D009474:Neurons / Q000166:cytology* / Q000648:ultrastructure; D009836:Oligodendroglia / Q000166:cytology* / Q000648:ultrastructure; D009924:Organ Culture Techniques; D011247:Pregnancy
D015415:Biomarkers