Cultures of cells from fetal rat brain: Methods to control composition, morphology, and biochemical activity
✍ Scribed by Melissa J. Mahoney; W. Mark Saltzman
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 458 KB
- Volume
- 62
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
Fetal tissue transplantation is a promising new approach for the treatment of neurodegenerative diseases, but the optimal conditions for preparing cells for transplantation have not been defined. The growth of a population of septal brain cells, primarily containing cholinergic neurons and glia, was characterized after seeding at densities from 5 × 10 4 to 6 × 10 5 cells/cm 2 , on polystyrene-, collagen-, laminin-, and fibronectin-coated surfaces, in the presence of serum and/or serum-free medium. Differentiated glial cells were selected by culture on fibronectin or laminin surfaces, in the presence of low amounts of serum (2.5% FBS) and G5, a soluble factor containing EGF and insulin. Differentiated neuronal cells were selected by culture on laminin, in the presence of low amounts of serum (2.5% FBS) and N2, a soluble factor containing supplemental hormones. In each case, a minimum seeding density of 1 × 10 5 cells/cm 2 was required. Neuronal growth could be maintained long term (21 days) with high levels of neuronal activity (ChAT activity).