Core\u2013shell hydrogel microcapsules enable formation of human pluripotent stem cell spheroids and their cultivation in a stirred bioreactor

Pouria Fattahi,Jadon Wolf,Ali Rahimian,Caden D Duffy,Quinn P Peterson,Kihak Gwon,Harihara Baskaran,Michael Q Slama,Alexander Revzin,Alan M Gonzalez-Suarez,Gulnaz Stybayeva

doi:10.1038/s41598-021-85786-2

Pouria Fattahi, Jadon Wolf + Show 9 more

Open Access

https://doi.org/10.1038/s41598-021-85786-2

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Journal: Scientific Reports	Publication Date: Mar 30, 2021
Citations: 32	License type: open-access

Affiliation: Mayo Clinic, Case Western Reserve University

Abstract

Cellular therapies based on human pluripotent stem cells (hPSCs) offer considerable promise for treating numerous diseases including diabetes and end stage liver failure. Stem cell spheroids may be cultured in stirred bioreactors to scale up cell production to cell numbers relevant for use in humans. Despite significant progress in bioreactor culture of stem cells, areas for improvement remain. In this study, we demonstrate that microfluidic encapsulation of hPSCs and formation of spheroids. A co-axial droplet microfluidic device was used to fabricate 400 μm diameter capsules with a poly(ethylene glycol) hydrogel shell and an aqueous core. Spheroid formation was demonstrated for three hPSC lines to highlight broad utility of this encapsulation technology. In-capsule differentiation of stem cell spheroids into pancreatic β-cells in suspension culture was also demonstrated.

Highlights

Cellular therapies based on human pluripotent stem cells offer considerable promise for treating numerous diseases including diabetes and end stage liver failure
Spheroid formation in standard suspension cultures occur due to random adhesive interactions between single cells inoculated into the bioreactor with spheroid assembly being a function of both cell concentration and the speed of rotation/agitation[9]
It is worth noting that that the tallest channel in the flow focusing microfluidic device is 300 μm which means that the resultant microparticles should have a diameter of ~ 300 μm and not 400 μm as mentioned above

Summary

Introduction

Cellular therapies based on human pluripotent stem cells (hPSCs) offer considerable promise for treating numerous diseases including diabetes and end stage liver failure. We employed a similar co-axial flow focusing microfluidic device to fabricate microcapsules with PEG hydrogel shell and aqueous core and demonstrated that non-proliferative primary hepatocytes assembled into spheroids upon encapsulation and remained functional for at least two weeks of culture[35]. We assessed the utility of the microcapsules with hydrogel shell and aqueous core for encapsulation of hPSCs (see Fig. 1A).

Results

Conclusion