Accéder au contenu
Merck

Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture.

Nature methods (2015-05-26)
Anca M Paşca, Steven A Sloan, Laura E Clarke, Yuan Tian, Christopher D Makinson, Nina Huber, Chul Hoon Kim, Jin-Young Park, Nancy A O'Rourke, Khoa D Nguyen, Stephen J Smith, John R Huguenard, Daniel H Geschwind, Ben A Barres, Sergiu P Paşca
RÉSUMÉ

The human cerebral cortex develops through an elaborate succession of cellular events that, when disrupted, can lead to neuropsychiatric disease. The ability to reprogram somatic cells into pluripotent cells that can be differentiated in vitro provides a unique opportunity to study normal and abnormal corticogenesis. Here, we present a simple and reproducible 3D culture approach for generating a laminated cerebral cortex-like structure, named human cortical spheroids (hCSs), from pluripotent stem cells. hCSs contain neurons from both deep and superficial cortical layers and map transcriptionally to in vivo fetal development. These neurons are electrophysiologically mature, display spontaneous activity, are surrounded by nonreactive astrocytes and form functional synapses. Experiments in acute hCS slices demonstrate that cortical neurons participate in network activity and produce complex synaptic events. These 3D cultures should allow a detailed interrogation of human cortical development, function and disease, and may prove a versatile platform for generating other neuronal and glial subtypes in vitro.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Bicarbonate de sodium, powder, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Albumine de sérum bovin, fatty acid free, low endotoxin, lyophilized powder, BioReagent, suitable for cell culture, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Chlorure de sodium, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
Chlorure de sodium solution, 0.9% in water, BioXtra, suitable for cell culture
Sigma-Aldrich
Chlorure de sodium, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
Chlorure de sodium solution, 5 M in H2O, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
L-cystéine, 97%
Sigma-Aldrich
Anticorps anti-NeuN, clone A60, clone A60, Chemicon®, from mouse
Sigma-Aldrich
L-cystéine, from non-animal source, BioReagent, suitable for cell culture, ≥98%
SAFC
Chlorure de sodium solution, 5 M
Sigma-Aldrich
Biocytine, ≥98% (TLC)
Sigma-Aldrich
L-cystéine, BioUltra, ≥98.5% (RT)
Sigma-Aldrich
Chlorure de sodium solution, BioUltra, for molecular biology, ~5 M in H2O
Sigma-Aldrich
Chlorure de sodium, BioUltra, for molecular biology, ≥99.5% (AT)
Sigma-Aldrich
Dorsomorphin, ≥98% (HPLC)
Sigma-Aldrich
Anticorps anti-transporteur vésiculaire du glutamate de type 1, serum, Chemicon®
SAFC
L-cystéine
Supelco
Retinol solution, 100 μg/mL ± 25% (Refer to COA) (Ethanol with 0.1% (w/v) BHT), ampule of 1 mL, reference material, Cerilliant®
Sigma-Aldrich
Earle′s Balanced Salt Solution 10x, Without sodium bicarbonate, 10 ×, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
L-cystéine, ≥97%, FG
Sigma-Aldrich
Chlorure de sodium, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
L-cystéine, produced by Wacker Chemie AG, Burghausen, Germany, ≥98.0%
Sigma-Aldrich
Sodium chloride-35Cl, 99 atom % 35Cl
Sigma-Aldrich
Bicarbonate de sodium, BioXtra, 99.5-100.5%
Sigma-Aldrich
Sodium bicarbonate-12C, 99.9 atom % 12C
Sigma-Aldrich
Chlorure de sodium solution, 0.85%
Sigma-Aldrich
Anti-Brn-2 (POU3F2) Antibody, clone 8C4.2, clone 8C4.2, from mouse