Isolation of neuronal precursors from differentiating P19 embryonal carcinoma cells by neuronal Tα1-promoter-driven GFP

Joon Kim, Seung Cheol Choi, Tae Hyung Kim, Ki Dong Kim, Si Young Cho, Sung Soo Park, Sang Ho Lee

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The induction of pluripotent P19 embryonal carcinoma (EC) cells with retinoic acid results in their differentiation into cells that resemble neurons, glia, and fibroblasts. To isolate and enrich the developing neurons from heterogeneously differentiating P19 EC cells, we used a recently introduced protocol combining the expression of green fluorescent protein (GFP) driven by a tissue-specific promoter and fluorescence-activated cell sorting. Cells were transfected with the gene for GFP, which is under the control of the neuronal Tα1 tubulin promoter. After four days of retinoic acid treatment, GFP was specifically detected in cells undergoing neuronal differentiation. Sorting of fluorescent differentiating P19 EC transfectants yielded populations highly enriched in neuronal precursors and neurons. Immunoreactivity for nestin and neurofilament was observed in 80 and 25% of the sorted cell population, respectively. These results demonstrate that differentiated neuronal precursor cells can be efficiently isolated from differentiating pluripotent embryonic cells in vitro, suggesting that this method can reproducibly provide homogeneous materials for further studies on neurogenesis.

Original languageEnglish
Pages (from-to)631-638
Number of pages8
JournalInternational Journal of Developmental Neuroscience
Volume19
Issue number7
DOIs
Publication statusPublished - 2001 Nov 17

Fingerprint

Embryonal Carcinoma Stem Cells
Green Fluorescent Proteins
Tretinoin
Neurons
Tubulin Modulators
Embryonal Carcinoma
Nestin
Intermediate Filaments
Neurogenesis
Neuroglia
Population
Cell Differentiation
Flow Cytometry
Fibroblasts
Genes

Keywords

  • Cell sorting
  • GFP
  • Neuronal differentiation
  • Neuronal precursor
  • P19 EC cell
  • Tα1 tubulin promoter

ASJC Scopus subject areas

  • Developmental Biology
  • Developmental Neuroscience

Cite this

Isolation of neuronal precursors from differentiating P19 embryonal carcinoma cells by neuronal Tα1-promoter-driven GFP. / Kim, Joon; Choi, Seung Cheol; Kim, Tae Hyung; Kim, Ki Dong; Cho, Si Young; Park, Sung Soo; Lee, Sang Ho.

In: International Journal of Developmental Neuroscience, Vol. 19, No. 7, 17.11.2001, p. 631-638.

Research output: Contribution to journalArticle

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