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  • HIPRO: A High-Efficiency, Hypoxia-Induced Protocol for Generation of Photoreceptors in Retinal Organoids from Mouse Pluripotent Stem Cells.

HIPRO: A High-Efficiency, Hypoxia-Induced Protocol for Generation of Photoreceptors in Retinal Organoids from Mouse Pluripotent Stem Cells.

STAR protocols (2020-08-06)
Holly Y Chen, Ryan A Kelley, Anand Swaroop
ABSTRACT

Mouse pluripotent stem cells can be efficiently differentiated into retinal organoids with polarized, laminated neural retina harboring all retinal cell types by the Hypoxia-Induced Generation of Photoreceptor in Retinal Organoids (HIPRO) protocol. In our recent publication, we modified the HIPRO protocol on the basis of comparative transcriptome analyses to facilitate photoreceptor biogenesis and maturation. Here, we provide a detailed protocol for efficient generation of retinal organoids from mouse pluripotent stem cells. For complete details on the use and execution of this protocol, please refer to (Chen et al., 2016, DiStefano et al., 2018, Brooks et al., 2019).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Brn-3a Antibody, POU-domain protein, clone 5A3.2, culture supernatant, clone 5A3.2, Chemicon®
Sigma-Aldrich
Anti-Protein Kinase Cα antibody produced in rabbit, whole antiserum
Sigma-Aldrich
Anti-Glutamine Synthetase Antibody, clone GS-6, clone GS-6, Chemicon®, from mouse
Sigma-Aldrich
Anti-Calbindin D-28K (Ab-1) Rabbit pAb, lyophilized, Calbiochem®