A novel human astrocyte cell line (A735) with astrocyte-specific neurotransmitter function
T N Price1, J F Burke, L V Mayne
1Trafford Centre for Graduate Medical Education and Research, Division of Molecular Medicine, University of Sussex, Falmer Brighton, United Kingdom.
In Vitro Cellular & Developmental Biology. Animal
|September 4, 1999
Summary
Researchers developed immortal human astrocyte cell lines using SV40 T antigen, overcoming challenges in brain cell research. These A735 cells retain key glial functions for studying brain physiology.
Area of Science:
- Neuroscience
- Cell Biology
- Genetics
Background:
- Establishing immortal human brain cell lines is difficult, hindering studies on brain cell function and physiology.
- Limited availability of reliable human glial cell lines restricts research progress.
Purpose of the Study:
- To develop immortal human astrocyte cell lines using recombinant DNA technology.
- To assess the functional properties and phenotype of SV40 T antigen-immortalized astrocytes.
Main Methods:
- Transfection of primary human astrocytes with SV40 T antigen expression vectors (pSV3neo or p735.6).
- Establishment and characterization of immortalized cell lines, including A735.
- Immunohistochemistry and Western blot analysis for astrocyte markers and function.
- Assay of neurotransmitter transport (GABA and glutamate).
Main Results:
- Successfully generated immortal human astrocyte cell lines with extended lifespans.
- The A735 cell line retained morphological and growth characteristics of type 1 astrocytes.
- Demonstrated differentiated but immature astrocyte phenotype and glial-specific neurotransmitter transport.
- SV40 T antigen-induced proliferation impacted glial fibrillary acidic protein expression.
Conclusions:
- SV40 T antigen effectively immortalizes human astrocytes while preserving key glial functions.
- A735 represents a novel, non-tumor-derived immortal human glial cell line.
- This advancement facilitates research into human brain cell physiology and disease.
