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, sis 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.
| Original language | English |
|---|---|
| Pages (from-to) | 209-221 |
| Number of pages | 13 |
| Journal | Journal of Molecular Neuroscience |
| Volume | 11 |
| Issue number | 3 |
| DOIs | |
| State | Published - Dec 1 1998 |
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Keywords
- Conditioned medium
- Dopaminergic neurons
- Gene expression analysis
- Immunocytochemistry
- Karotype
- Neurodegenerative diseases
- Neuroprotection
- Parkinson's disease
- Tissue culture
- Ventral mescencephalon
ASJC Scopus subject areas
- Neuroscience(all)
- Biochemistry
- Genetics
Cite this
An immortalized, type-1 astrocyte of mesencephalic origin source of a dopaminergic neurotrophic factor. / Panchision, David M.; Martin-DeLeon, Patricia A.; Takeshima, Takao; Johnston, Jane M.; Shimoda, Kotaro; Tsoulfas, Pantelis; McKay, Ronald D G; Commissiong, John W.
In: Journal of Molecular Neuroscience, Vol. 11, No. 3, 01.12.1998, p. 209-221.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - An immortalized, type-1 astrocyte of mesencephalic origin source of a dopaminergic neurotrophic factor
AU - Panchision, David M.
AU - Martin-DeLeon, Patricia A.
AU - Takeshima, Takao
AU - Johnston, Jane M.
AU - Shimoda, Kotaro
AU - Tsoulfas, Pantelis
AU - McKay, Ronald D G
AU - Commissiong, John W.
PY - 1998/12/1
Y1 - 1998/12/1
N2 - 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, sis 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.
AB - 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, sis 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.
KW - Conditioned medium
KW - Dopaminergic neurons
KW - Gene expression analysis
KW - Immunocytochemistry
KW - Karotype
KW - Neurodegenerative diseases
KW - Neuroprotection
KW - Parkinson's disease
KW - Tissue culture
KW - Ventral mescencephalon
UR - http://www.scopus.com/inward/record.url?scp=0032432604&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032432604&partnerID=8YFLogxK
U2 - 10.1385/JMN:11:3:209
DO - 10.1385/JMN:11:3:209
M3 - Article
C2 - 10344791
AN - SCOPUS:0032432604
VL - 11
SP - 209
EP - 221
JO - Journal of Molecular Neuroscience
JF - Journal of Molecular Neuroscience
SN - 0895-8696
IS - 3
ER -