Biotechnol J. 2018 Aug 29:e1800323. doi: 10.1002/biot.201800323. [Epub ahead of print]

Long-term Retinal Differentiation of Human Induced Pluripotent Stem Cells in a Continuously Perfused Microfluidic Culture Device.

Abdolvand N¹, Tostoes R¹, Raimes W¹, Kumar V¹, Szita N¹, Veraitch F¹.

Abstract

BACKGROUND:

Understanding how microenvironmental cues influence cellular behaviour will enable development of efficient and robust pluripotent stem cell differentiation protocols. Unlike traditional cell culture dishes, microfluidic bioreactors can provide stable microenvironmental conditions by continuous medium perfusion at a controlled rate. The aim of this study was to investigate whether a microfluidic culture device could be used as a perfused platform for long-term cell culture processes such as the retinal differentiation of human induced pluripotent stem cells.

METHODS AND RESULTS:

The perfusion flow rate was established based on the degradation and consumption of growth factors (DKK-1, Noggin, IGF-1 and bFGF) and utilising the Péclet number. The device's performance analysed by qRT-PCR showed improvements compared to the well-plate control as characterised by significantly higher expression of the markers Pax 6, Chx10 and Crx on day 5, Nrl on day 10, Crx and Rhodopsin on day 21.

CONCLUSIONS:

Optimisation of perfusion rate is an important operating variable in development of robust processes for differentiation cultures. Result demonstrates convective delivery of nutrients via perfusion has a significant impact upon the expression of key retinal markers. Novelty: This study was the first continuously perfused long-term (21 days) retinal differentiation of hiPSCs in a microfluidic device.

This article is protected by copyright. All rights reserved.