A scalable and tunable thermoreversible polymer for 3D human pluripotent stem cell biomanufacturing

Hunter J Johnson; Saheli Chakraborty; Riya J Muckom; Nitash P Balsara; David V Schaffer

doi:10.1016/j.isci.2022.104971

A scalable and tunable thermoreversible polymer for 3D human pluripotent stem cell biomanufacturing

iScience. 2022 Aug 25;25(10):104971. doi: 10.1016/j.isci.2022.104971. eCollection 2022 Oct 21.

Authors

Hunter J Johnson^{1

2}, Saheli Chakraborty³, Riya J Muckom⁴, Nitash P Balsara^{3

4

5

6}, David V Schaffer^{1

4

7

8}

Affiliations

¹ Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA.
² Graduate Program in Bioengineering, University of California, California, San Francisco and University of California, Berkeley, Berkeley, CA 94720, USA.
³ Energy Storage & Distribution Resources Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
⁴ Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA 94720, USA.
⁵ Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
⁶ Joint Center for Energy Storage Research, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
⁷ Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, CA 94720, USA.
⁸ Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.

Abstract

Human pluripotent stem cells (hPSCs) are an exciting and promising source to enable cell replacement therapies for a variety of unmet medical needs. Though hPSCs can be successfully derived into numerous physiologically relevant cell types, effective translation to the clinic is limited by challenges in scalable production of high-quality cells, cellular immaturity following the differentiation process, and the use of animal-derived components in culture. To address these limitations, we have developed a fully defined, reproducible, and tunable thermoreversible polymer for high-quality, scalable 3D cell production. Our reproducible synthesis method enables precise control of gelation temperature (24°C-32°C), hydrogel stiffness (100-4000 Pa), and the prevention of any unintended covalent crosslinking. After material optimization, we demonstrated hPSC expansion, pluripotency maintenance, and differentiation into numerous lineages within the hydrogel. Overall, this 3D thermoreversible hydrogel platform has broad applications in scalable, high-quality cell production to overcome the biomanufacturing burden of stem cell therapy.

Keywords: Biotechnology; Cell biology; Materials in biotechnology; Stem cells research.