Three to four-year-old nonpassaged EGF-responsive neural progenitor cells

Proliferation, apoptosis, and DNA repair

F. C. Zhou, M. R. Kelley, Y. H. Chiang, P. Young

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Epidermal growth factor responsive (EGFr) neural progenitor (NP) cells have been shown to be a potential alternative tissue source for neural transplantation and for developmental study. We have shown that nonpassaged EGFr NP cells can self-renew for 2 years in neurospheres and can robustly differentiate into glia and a number of neuronal cell types. We are now attempting to investigate if the EGFr NP cells will die or continue to live beyond the life span of the donor. In addition, we and other investigators have also found that EGFr NP cells, after transplant, retain only a small number of cells in the transplant site. In this study, we investigate the plasticity and fate of the EGFr NP cells. Using the nonpassaged method, we found EGFr NP cells live in the EGF supplement medium for over 4 years - the longest-lived EGFr NP cells ever reported. The 4-year-old striatal or cortical EGFr neurospheres, when subplated with substrate coating, migrate out of neurospheres and have robust growth with many processes. Furthermore, when nucleotide marker bromodeoxyuridine (BrdU) was added 3 days prior to the subplating, the EGFr NP cells were labeled positively with BrdU in the nucleus, indicating active proliferation activity. Meanwhile two other events were also found in the long-term EGFr NP cells. In the midst of the proliferation, apoptosis occurred. A subpopulation of EGFr NP cells are undergoing programmed cell death as indicated by the cell morphology and the TUNEL staining for DNA strand breaks. The TUNEL fluorescein-staining indicates that over 50% of EGFr NP cells are positive in the nuclei. On the other hand, we have also found that the major base excision repair enzyme, APE/ref-1, which is responsible for recognizing and repairing baseless sites in DNA, was present in the progenitor cells. However, in those cells undergoing apoptosis, APE/ref-1 levels were dramatically reduced or missing, and only a small percentage of cells were TUNEL and APE/ref-1 positive. These observations indicate that EGFr neural progenitor cells can live beyond the life span of the donor animal. The longevity of these cells in culture may be enhanced due to decreased apoptosis and the retention of normal DNA repair capacity. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)200-208
Number of pages9
JournalExperimental Neurology
Volume164
Issue number1
DOIs
Publication statusPublished - Jul 2000
Externally publishedYes

Fingerprint

Epidermal Growth Factor
DNA Repair
Stem Cells
Cell Proliferation
Apoptosis
In Situ Nick-End Labeling
Bromodeoxyuridine
Staining and Labeling
Transplants
Corpus Striatum
DNA Breaks
Fluorescein
Neuroglia
Cell Death
Nucleotides
Cell Culture Techniques
Cell Count
Transplantation
Research Personnel

Keywords

  • APE/ref-1
  • Epidermal growth factor
  • Neural stem cells
  • Programmed cell death
  • TUNEL

ASJC Scopus subject areas

  • Neurology
  • Neuroscience(all)

Cite this

Three to four-year-old nonpassaged EGF-responsive neural progenitor cells : Proliferation, apoptosis, and DNA repair. / Zhou, F. C.; Kelley, M. R.; Chiang, Y. H.; Young, P.

In: Experimental Neurology, Vol. 164, No. 1, 07.2000, p. 200-208.

Research output: Contribution to journalArticle

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