TY - JOUR
T1 - Bioactive hydrogel microcapsules for guiding stem cell fate decisions by release and reloading of growth factors
AU - Gwon, Kihak
AU - Hong, Hye Jin
AU - Gonzalez-Suarez, Alan M.
AU - Slama, Michael Q.
AU - Choi, Daheui
AU - Hong, Jinkee
AU - Baskaran, Harihara
AU - Stybayeva, Gulnaz
AU - Peterson, Quinn P.
AU - Revzin, Alexander
N1 - Publisher Copyright:
© 2021 The Authors
PY - 2022/9
Y1 - 2022/9
N2 - Human pluripotent stem cells (hPSC) hold considerable promise as a source of adult cells for treatment of diseases ranging from diabetes to liver failure. Some of the challenges that limit the clinical/translational impact of hPSCs are high cost and difficulty in scaling-up of existing differentiation protocols. In this paper, we sought to address these challenges through the development of bioactive microcapsules. A co-axial flow focusing microfluidic device was used to encapsulate hPSCs in microcapsules comprised of an aqueous core and a hydrogel shell. Importantly, the shell contained heparin moieties for growth factor (GF) binding and release. The aqueous core enabled rapid aggregation of hPSCs into 3D spheroids while the bioactive hydrogel shell was used to load inductive cues driving pluripotency maintenance and endodermal differentiation. Specifically, we demonstrated that one-time, 1 h long loading of pluripotency signals, fibroblast growth factor (FGF)-2 and transforming growth factor (TGF)-β1, into bioactive microcapsules was sufficient to induce and maintain pluripotency of hPSCs over the course of 5 days at levels similar to or better than a standard protocol with soluble GFs. Furthermore, stem cell-carrying microcapsules that previously contained pluripotency signals could be reloaded with an endodermal cue, Nodal, resulting in higher levels of endodermal markers compared to stem cells differentiated in a standard protocol. Overall, bioactive heparin-containing core-shell microcapsules decreased GF usage five-fold while improving stem cell phenotype and are well suited for 3D cultivation of hPSCs.
AB - Human pluripotent stem cells (hPSC) hold considerable promise as a source of adult cells for treatment of diseases ranging from diabetes to liver failure. Some of the challenges that limit the clinical/translational impact of hPSCs are high cost and difficulty in scaling-up of existing differentiation protocols. In this paper, we sought to address these challenges through the development of bioactive microcapsules. A co-axial flow focusing microfluidic device was used to encapsulate hPSCs in microcapsules comprised of an aqueous core and a hydrogel shell. Importantly, the shell contained heparin moieties for growth factor (GF) binding and release. The aqueous core enabled rapid aggregation of hPSCs into 3D spheroids while the bioactive hydrogel shell was used to load inductive cues driving pluripotency maintenance and endodermal differentiation. Specifically, we demonstrated that one-time, 1 h long loading of pluripotency signals, fibroblast growth factor (FGF)-2 and transforming growth factor (TGF)-β1, into bioactive microcapsules was sufficient to induce and maintain pluripotency of hPSCs over the course of 5 days at levels similar to or better than a standard protocol with soluble GFs. Furthermore, stem cell-carrying microcapsules that previously contained pluripotency signals could be reloaded with an endodermal cue, Nodal, resulting in higher levels of endodermal markers compared to stem cells differentiated in a standard protocol. Overall, bioactive heparin-containing core-shell microcapsules decreased GF usage five-fold while improving stem cell phenotype and are well suited for 3D cultivation of hPSCs.
KW - 3D stem cell culture
KW - Bioactive core-shell microcapsule
KW - Droplet microfluidics
KW - Growth factor release
KW - Stem cell differentiation
UR - http://www.scopus.com/inward/record.url?scp=85121672262&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85121672262&partnerID=8YFLogxK
U2 - 10.1016/j.bioactmat.2021.12.008
DO - 10.1016/j.bioactmat.2021.12.008
M3 - Article
AN - SCOPUS:85121672262
SN - 2452-199X
VL - 15
SP - 1
EP - 14
JO - Bioactive Materials
JF - Bioactive Materials
ER -