PMID- 10446410
OWN - NLM
STAT- MEDLINE
DCOM- 19991012
LR  - 20191103
IS  - 1385-299X (Print)
IS  - 1385-299X (Linking)
VI  - 4
IP  - 2
DP  - 1999 Jul
TI  - An improved method for dissociation and aggregate culture of human fetal brain 
      cells in serum-free medium.
PG  - 156-64
AB  - 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.] [16], 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 which expression
FAU - Barnea, A
AU  - Barnea A
AD  - Department of Obstetrics and Gynecology, The University of Texas Southwestern 
      Medical Center at Dallas, Dallas, TX 75235-9032, USA. 
      ayalla.barnea@email.swmed.edu
FAU - Roberts, J
AU  - Roberts J
LA  - eng
GR  - NS32207/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Netherlands
TA  - Brain Res Brain Res Protoc
JT  - Brain research. Brain research protocols
JID - 9716650
RN  - 0 (Culture Media, Serum-Free)
RN  - 0 (Neuropeptide Y)
RN  - 51110-01-1 (Somatostatin)
RN  - EC 3.1.- (Deoxyribonucleases)
RN  - EC 3.4.21.4 (Trypsin)
SB  - IM
MH  - Brain/*cytology/embryology
MH  - Cell Aggregation
MH  - Cell Culture Techniques/*methods
MH  - Cell Separation/*methods
MH  - Cells, Cultured
MH  - Culture Media, Serum-Free
MH  - Deoxyribonucleases
MH  - Fetus/*cytology
MH  - Humans
MH  - Neuroglia/cytology
MH  - Neurons/cytology
MH  - Neuropeptide Y/biosynthesis
MH  - Somatostatin/biosynthesis
MH  - Specimen Handling
MH  - Stress, Mechanical
MH  - Trypsin
EDAT- 1999/08/14 00:00
MHDA- 1999/08/14 00:01
CRDT- 1999/08/14 00:00
PHST- 1999/08/14 00:00 [pubmed]
PHST- 1999/08/14 00:01 [medline]
PHST- 1999/08/14 00:00 [entrez]
AID - S1385-299X(99)00015-X [pii]
AID - 10.1016/s1385-299x(99)00015-x [doi]
PST - ppublish
SO  - Brain Res Brain Res Protoc. 1999 Jul;4(2):156-64. doi: 
      10.1016/s1385-299x(99)00015-x.