Human Primary Mixed Brain Cultures: Preparation, Differentiation, Characterization and Application to Neuroscience Research

Overview

Journal Mol Brain

Publisher Biomed Central

Specialties Molecular Biology
Neurology

Date 2014 Sep 17

PMID 25223359

Citations 25

Authors

Balmiki Ray

Nipun Chopra

Justin M Long

Debomoy K Lahiri

Affiliations

Soon will be listed here.

Abstract

Background: Culturing primary cortical neurons is an essential neuroscience technique. However, most cultures are derived from rodent brains and standard protocols for human brain cultures are sparse. Herein, we describe preparation, maintenance and major characteristics of a primary human mixed brain culture, including neurons, obtained from legally aborted fetal brain tissue. This approach employs standard materials and techniques used in the preparation of rodent neuron cultures, with critical modifications.

Results: This culture has distinct differences from rodent cultures. Specifically, a significant numbers of cells in the human culture are derived from progenitor cells, and the yield and survival of the cells grossly depend on the presence of bFGF. In the presence of bFGF, this culture can be maintained for an extended period. Abundant productions of amyloid-β, tau and proteins make this a powerful model for Alzheimer's research. The culture also produces glia and different sub-types of neurons.

Conclusion: We provide a well-characterized methodology for human mixed brain cultures useful to test therapeutic agents under various conditions, and to carry forward mechanistic and translational studies for several brain disorders.

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References

GEIGER R . Subcultures of adult mammalian brain cortex in vitro. Exp Cell Res. 1958; 14(3):541-66. DOI: 10.1016/0014-4827(58)90160-5. View

Ray B, Bailey J, Sarkar S, Lahiri D . Molecular and immunocytochemical characterization of primary neuronal cultures from adult rat brain: Differential expression of neuronal and glial protein markers. J Neurosci Methods. 2009; 184(2):294-302. PMC: 2892104. DOI: 10.1016/j.jneumeth.2009.08.018. View

Long J, Ray B, Lahiri D . MicroRNA-153 physiologically inhibits expression of amyloid-β precursor protein in cultured human fetal brain cells and is dysregulated in a subset of Alzheimer disease patients. J Biol Chem. 2012; 287(37):31298-310. PMC: 3438960. DOI: 10.1074/jbc.M112.366336. View

Brewer G, Torricelli J . Isolation and culture of adult neurons and neurospheres. Nat Protoc. 2007; 2(6):1490-8. DOI: 10.1038/nprot.2007.207. View

Bailey J, Lahiri D . A novel effect of rivastigmine on pre-synaptic proteins and neuronal viability in a neurodegeneration model of fetal rat primary cortical cultures and its implication in Alzheimer's disease. J Neurochem. 2009; 112(4):843-53. PMC: 2881332. DOI: 10.1111/j.1471-4159.2009.06490.x. View

Yu S, Zhang J, Zhao C, Zhang H, Xu Q . Isolation and characterization of the CD133+ precursors from the ventricular zone of human fetal brain by magnetic affinity cell sorting. Biotechnol Lett. 2004; 26(14):1131-6. DOI: 10.1023/B:BILE.0000035484.64499.ac. View

Ferrari D, Binda E, De Filippis L, Vescovi A . Isolation of neural stem cells from neural tissues using the neurosphere technique. Curr Protoc Stem Cell Biol. 2010; Chapter 2:Unit2D.6. DOI: 10.1002/9780470151808.sc02d06s15. View

Jhamandas J, Li Z, Westaway D, Yang J, Jassar S, MacTavish D . Actions of β-amyloid protein on human neurons are expressed through the amylin receptor. Am J Pathol. 2011; 178(1):140-9. PMC: 3070588. DOI: 10.1016/j.ajpath.2010.11.022. View

Mattson M, Rychlik B . Cell culture of cryopreserved human fetal cerebral cortical and hippocampal neurons: neuronal development and responses to trophic factors. Brain Res. 1990; 522(2):204-14. DOI: 10.1016/0006-8993(90)91462-p. View

10.

Bystron I, Blakemore C, Rakic P . Development of the human cerebral cortex: Boulder Committee revisited. Nat Rev Neurosci. 2008; 9(2):110-22. DOI: 10.1038/nrn2252. View

11.

Darbinyan A, Kaminski R, White M, Darbinian N, Khalili K . Isolation and propagation of primary human and rodent embryonic neural progenitor cells and cortical neurons. Methods Mol Biol. 2013; 1078:45-54. PMC: 4110112. DOI: 10.1007/978-1-62703-640-5_5. View

12.

Rieske P, Azizi S, Augelli B, Gaughan J, Krynska B . A population of human brain parenchymal cells express markers of glial, neuronal and early neural cells and differentiate into cells of neuronal and glial lineages. Eur J Neurosci. 2007; 25(1):31-7. DOI: 10.1111/j.1460-9568.2006.05254.x. View

13.

Csoregh L, Andersson E, Fried G . Transcriptional analysis of estrogen effects in human embryonic neurons and glial cells. Neuroendocrinology. 2008; 89(2):171-86. DOI: 10.1159/000153899. View

14.

Silani V, Pizzuti A, Strada O, Falini A, Buscaglia M, Scarlato G . Human neuronal cell viability demonstrated in culture after cryopreservation. Brain Res. 1988; 473(1):169-74. DOI: 10.1016/0006-8993(88)90331-9. View

15.

Vaccarino F, Schwartz M, Raballo R, Nilsen J, Rhee J, Zhou M . Changes in cerebral cortex size are governed by fibroblast growth factor during embryogenesis. Nat Neurosci. 1999; 2(3):246-53. DOI: 10.1038/6350. View

16.

Liesi P, Fried G, Stewart R . Neurons and glial cells of the embryonic human brain and spinal cord express multiple and distinct isoforms of laminin. J Neurosci Res. 2001; 64(2):144-67. DOI: 10.1002/jnr.1061. View

17.

Leblanc A . Increased production of 4 kDa amyloid beta peptide in serum deprived human primary neuron cultures: possible involvement of apoptosis. J Neurosci. 1995; 15(12):7837-46. PMC: 6577962. View

18.

Negishi T, Ishii Y, Kyuwa S, Kuroda Y, Yoshikawa Y . Primary culture of cortical neurons, type-1 astrocytes, and microglial cells from cynomolgus monkey (Macaca fascicularis) fetuses. J Neurosci Methods. 2003; 131(1-2):133-40. DOI: 10.1016/j.jneumeth.2003.08.006. View

19.

Grynkiewicz G, Poenie M, Tsien R . A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985; 260(6):3440-50. View

20.

Roediger B, Armati P . Oxidative stress induces axonal beading in cultured human brain tissue. Neurobiol Dis. 2003; 13(3):222-9. DOI: 10.1016/s0969-9961(03)00038-x. View