Journal of Molecular Neuroscience

, Volume 11, Issue 3, pp 209–221 | Cite as

An immortalized, type-1 astrocyte of mescencephalic origin source of a dopaminergic neurotrophic factor

  • David M. Panchision
  • Patricia A. Martin-DeLeon
  • Takao Takeshima
  • Jane M. Johnston
  • Kotaro Shimoda
  • Pantelis Tsoulfas
  • Ronald D. G. McKay
  • John W. Commissiong
Article

Abstract

Rat embryonic d 14 (E14) mesencephalic cells, 2.5% of which are glioblasts, were incubated in medium containing 10% of fetal bovine serum for 12 h and subsequently expanded in a serum-free medium using basic fibroblast growth factor (bFGF) as the mitogen. On a single occasion, after more than 15 d in culture, several islets of proliferating, glial-like cells were observed in one dish. The cells, when isolated and passaged, proliferated rapidly in either a serum-free or serum-containing growth medium. Subsequent immunocytochemical analysis showed that they stained positive for GFAP and vimentin, and negative for A2B5, O4, GalC, and MAP2. Serum-free conditioned medium (CM) prepared from these cells caused a fivefold increase in survival and promoted neuritic expansion of E14 mesencephalic dopaminergic neurons in culture. These actions are similar to those exerted by CM derived from primary, mesencephalic type-1 astrocytes. The pattern of expression of the region-selective genes; wnt-1, en-1, showed that 70% of the cells were heteroploid, and of these, 50% were tetraploid. No apparent decline in proliferative capacity has been observed after 25 passages. The properties of this cell line, named ventral mesencephalic cell line one (VMCL1), are consistent with those of an immortalized, type-1 astrocyte. The mesencephalic origin of the cell line, and the pattern and potency of the neurotrophic activity exerted by the CM, strongly suggest that the neurotrophic factor(s) identified are novel, and will likely be strong candidates with clinical utility for the treatment of Parkinson’s disease.

Index Entries

Conditioned medium dopaminergic neurons gene expression analysis immunocytochemistry karotype neurodegenerative diseases neuroprotection Parkinson’s disease tissue culture ventral mescencephalon 

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Copyright information

© Humana Press Inc 1999

Authors and Affiliations

  • David M. Panchision
    • 1
  • Patricia A. Martin-DeLeon
    • 2
  • Takao Takeshima
    • 3
  • Jane M. Johnston
    • 4
  • Kotaro Shimoda
    • 5
  • Pantelis Tsoulfas
    • 1
  • Ronald D. G. McKay
    • 1
  • John W. Commissiong
    • 1
  1. 1.National Institute of Neurological Disorders and StrokeNational Institutes of HealthBethesda
  2. 2.Division of Molecular Biology, Department of Biological SciencesUniversity of DelawareNewark
  3. 3.Division of Neurology, Institute of Neurological SciencesTottori University School of MedicineYonagoJapan
  4. 4.Department of Neurological Surgery, Henry and Lucy Moses Division, Montefiore Medical CenterUniversity Hospital for the Albert Einstein College of MedicineBronx
  5. 5.Division of NeurologyNational Nishi Tottori HospitalTottoriJapan

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