Purification of cell populations from human fetal brain using flow cytometric techniques

R. Rozental, D. Gebhard, C. Padin, M. Urban, J. Y. Wu, D. C. Spray, F. C. Chiu

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

We recently established primary cultures from dissociated second trimester human fetal brains using a novel spin seeding method and characterized cellular populations with distinct phenotypes in these cultures. Here, we report that these neural cultures can be dissociated to single-cell suspensions, sorted by size using flow cytometry and re-seeded to yield cultures selectively enriched for the neuronal and glial cell populations. Sorted neurons were highly homogeneous, viable and extended processes by one day after re-seeding. These neurons expressed immunoreactivity for neurofilament protein, retained their GABAergic phenotype and were electrically excitable. Re-seeded astrocytes proliferated in culture and expressed glial fibrillary acidic protein. We describe the conditions required for the flow cytometric sorting and tissue culture assays as well as the morphological, immunocytochemical and electrophysiological characteristics of the sorted neuronal population.

Original languageEnglish (US)
Pages (from-to)161-170
Number of pages10
JournalDevelopmental Brain Research
Volume85
Issue number2
DOIs
StatePublished - Apr 18 1995

Keywords

  • Flow cytometry
  • Glutamic acid decarboxylase
  • Human neuron
  • Neurofilament protein

ASJC Scopus subject areas

  • Developmental Neuroscience
  • Developmental Biology

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