Brain Res Brain Res Protoc. 1999 Jul;4(2):156-64. Unique Identifier :
Moscona, in the early sixties [A.A. Moscona, Recombination of
dissociated cells and the development of cell aggregates, in: B.M.
Willmer (Ed.), Cells and Tissues in Culture, Academic Press, New York,
1965, pp. 489-529.] , discovered that aggregation of dissociated
cells is a property of embryonal cells. Several features of the
aggregate culture system are particularly attractive for the conduct of
biochemical and molecular studies on the human fetal brain. (i) All the
pertinent procedural parameters can be readily controlled and
standardized, resulting in a consistently reproducible system suitable
for quantitative analyses. (ii) Neuronal enriched aggregates can be
readily obtained, with minimal neurotoxicity. (iii) Aggregates can be
easily harvested for biochemical and molecular studies. Aggregate
cultures, generated from rodent fetal brains, have been extensively
utilized as a tool to study regulation of aminergic neurons [P.
Honegger, E. Richelson, Biochemical differentiation of mechanically
dissociated brain in aggregating cell culture, Brain Res. 109 (1976)
335-354; P. Honegger, E. Richelson, Biochemical differentiation of
aggregating cell cultures of different fetal rat brain regions, Brain
Res. 133 (1977) 329-339.] [11,12] and peptidergic neurons (neuropeptide
Y (NPY) and somatostatin (SRIF) [A. Barnea, E. Anthony, G. Lu, G. Cho,
Morphological differentiation of neuropeptide Y neurons in aggregate
cultures of dissociated fetal cortical cells: a model system for
glia-neuron paracrine interactions, Brain Res. 625 (1993) 313-322; A.
Barnea, G. Cho, G. Lu, M. Mathis, Brain-derived neurotrophic factor
induces functional expression and phenotypic differentiation of cultured
fetal neuropeptide Y producing neurons, J. Neurosci. Res. 42 (1995)
638-647; A. Barnea, A. Hajibeigi, G. Cho, P. Magni, Regulated production
and secretion of immunoreactive neuropeptide Y by aggregating fetal
brain cells in culture, Neuroendocrinology 54 (1991) 7-13; P. Magni, A.
Barnea, Forskolin and phorbol ester stimulation of neuropeptide Y (NPY)
production and secretion by aggregating fetal brain cells in culture:
evidence for regulation of NPY biosynthesis at transcriptional and
posttranscriptional levels, Endocrinology 130 (1992) 976-984.])
[4-6,14]. However, very few studies have utilized this system to study
regulatory processes of human fetal neurons/glia [M. McCarthy, L.
Resnik, F. Taub, R.V. Stewart, R.D. Dix, Infection of human neural cell
aggregate cultures with a clinical isolate of cytomegalovirus, J.
Neuropathol. Exp. Neurol. 50 (1991) 441-450; L. Pulliam, M.E. Berens,
M.L. Rosenblum, A normal human brain cell aggregate model for
neurobiological studies, J. Neurosci. Res. 21 (1988) 521-530.] [15,17].
In a series of studies in our laboratory [N. Aguila-Mansilla, A. Barnea,
Human fetal brain cells in aggregate culture: a model system to study
regulatory processes of the developing human neuropeptide Y (NPY)
producing neuron, Int. J. Dev. Neurosci. 14 (1996) 531-539; A. Barnea,
N. Aguila-Mansilla, H.T. Chute, A.A. Welcher, Comparison of neurotrophin
regulation of human and rat neuropeptide Y (NPY) neurons: induction of
NPY production in aggregate cultures derived from rat but not from human
fetal brains, Brain Res. 732 (1996) 52-60; A. Barnea, N.
Aguila-Mansilla, G. Lu, R.H. Ho, Opposite effects of astrocyte-derived
soluble factor(s) on the functional expression of fetal peptidergic
neurons in aggregate cultures: enhancement of neuropeptide Y and
suppression of somatostatin, J. Neurosci. Res. 50 (1997) 605-617; A.
Barnea, J. Roberts, R.H. Ho, Evidence for a synergistic effect of the
HIV-1 envelope protein gp120 and brain-derived neurotrophic factor
(BDNF) leading to enhanced expression of somatostatin neurons in
aggregate cultures derived from the human fetal cortex, Brain Res. 815
(1999) 349-357.] [1-3,7], we have established a human-derived aggregate
culture system, maintained in serum-free medium for up to 28 days, in
JOURNAL ARTICLE Brain/*CYTOLOGY/EMBRYOLOGY Cell Aggregation Cell
Culture/*METHODS Cell Separation/*METHODS Cells, Cultured Culture
Media, Serum-Free Deoxyribonucleases Fetus/*CYTOLOGY Human
Neuroglia/CYTOLOGY Neurons/CYTOLOGY Neuropeptide Y/BIOSYNTHESIS
Somatostatin/BIOSYNTHESIS Specimen Handling Stress, Mechanical
Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S. Trypsin