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

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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.

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Author notes

  1. Pantelis Tsoulfas

    Present address: The Miami Project, University of Miami School of Medicine, 33136, Miami, FL

  2. John W. Commissiong

    Present address: Neurotrophic Research Corp., Suite 100, 8200 Wisconsin Avenue, 20814, Bethesda, MD

Authors and Affiliations

  1. National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bldg. 36/3D02, 36 Convent Drive MSC 4092, 20892-4092, Bethesda, MD

    David M. Panchision, Pantelis Tsoulfas, Ronald D. G. McKay & John W. Commissiong

  2. Division of Molecular Biology, Department of Biological Sciences, University of Delaware, 19716, Newark, DE

    Patricia A. Martin-DeLeon

  3. Division of Neurology, Institute of Neurological Sciences, Tottori University School of Medicine, 86 Nishimachi, 683, Yonago, Japan

    Takao Takeshima

  4. Department of Neurological Surgery, Henry and Lucy Moses Division, Montefiore Medical Center, University Hospital for the Albert Einstein College of Medicine, 111 East 210th Street, 10467-2490, Bronx, NY

    Jane M. Johnston

  5. Division of Neurology, National Nishi Tottori Hospital, Mitsu 876, 68902, Tottori, Japan

    Kotaro Shimoda

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  1. David M. Panchision
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Panchision, D.M., Martin-DeLeon, P.A., Takeshima, T. et al. An immortalized, type-1 astrocyte of mescencephalic origin source of a dopaminergic neurotrophic factor. J Mol Neurosci 11, 209–221 (1998). https://doi.org/10.1385/JMN:11:3:209

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