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Cultures of cells from fetal rat brain: Methods to control composition, morphology, and biochemical activity
Author(s) -
Mahoney Melissa J.,
Saltzman W. Mark
Publication year - 1999
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19990220)62:4<461::aid-bit9>3.0.co;2-a
Subject(s) - laminin , fibronectin , fetal bovine serum , transplantation , microbiology and biotechnology , fetal tissue transplantation , chemically defined medium , biology , extracellular matrix , cell culture , epidermal growth factor , chemistry , fetus , population , biochemistry , medicine , cell , in vitro , pregnancy , genetics , environmental health
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). © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 62: 461–467, 1999.