Early retinal differentiation of human pluripotent stem cells in microwell suspension cultures

Objective To develop a microwell suspension platform for the adaption of attached stem cell differentiation protocols into mixed suspension culture. Results We adapted an adherent protocol for the retinal differentiation of human induced pluripotent stem cells (hiPSCs) using a two-step protocol. Est...

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Bibliographic Details
Published in:Biotechnology letters 2017-02, Vol.39 (2), p.339-350
Main Authors: Sharma, Vishal S., Khalife, Rana, Tostoes, Rui, Leung, Leonard, Kinsella, Rose, Ruban, Ludmilla, Veraitch, Farlan S.
Format: Article
Language:English
Subjects:
Applied Microbiology
Biochemistry
Biomedical and Life Sciences
Bioreactors
Biotechnology
Cell Culture Techniques
Cell differentiation
Cell Differentiation - physiology
Culture
Differentiation
Embryoid Bodies - cytology
Humans
Induced Pluripotent Stem Cells - cytology
Life Sciences
Markers
Membrane reactors
Microbiology
Optimization
Original Research Paper
Pluripotent Stem Cells - cytology
Protocol
Retina - cytology
Shaking
Stem cells
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Summary:Objective To develop a microwell suspension platform for the adaption of attached stem cell differentiation protocols into mixed suspension culture. Results We adapted an adherent protocol for the retinal differentiation of human induced pluripotent stem cells (hiPSCs) using a two-step protocol. Establishing the optimum embryoid body (EB) starting size and shaking speed resulted in the translation of the original adherent process into suspension culture. Embryoid bodies expanded in size as the culture progressed resulting in the expression of characteristic markers of early (Rx, Six and Otx2) and late (Crx, Nrl and Rhodopsin) retinal differentiation. The new process also eliminated the use of matrigel, an animal-derived extracellular matrix coating. Conclusions Shaking microwells offer a fast and cost-effective method for proof-of-concept studies to establish whether pluripotent stem cell differentiation processes can be translated into mixed suspension culture.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-016-2244-7