TY - JOUR
T1 - An integrated biomanufacturing platform for the large-scale expansion and neuronal differentiation of human pluripotent stem cell-derived neural progenitor cells
AU - Srinivasan, Gayathri
AU - Morgan, Daylin
AU - Varun, Divya
AU - Brookhouser, Nicholas
AU - Brafman, David
N1 - Funding Information:
Funding for this work was provided by the NIH-NIBIB (5R21EB020767) and the Arizona Biomedical Research Commission. N.B. was supported by a fellowship from the International Foundation for Ethical Research. We would like to thank Jing Zhao and Guojun Bu for assistance with the astrocyte differentiation protocols.
Funding Information:
Funding for this work was provided by the NIH - NIBIB (5R21EB020767) and the Arizona Biomedical Research Commission . N.B. was supported by a fellowship from the International Foundation for Ethical Research . We would like to thank Jing Zhao and Guojun Bu for assistance with the astrocyte differentiation protocols.
Publisher Copyright:
© 2018 Acta Materialia Inc.
PY - 2018/7/1
Y1 - 2018/7/1
N2 - Human pluripotent stem cell derived neural progenitor cells (hNPCs) have the unique properties of long-term in vitro expansion as well as differentiation into the various neurons and supporting cell types of the central nervous system (CNS). Because of these characteristics, hNPCs have tremendous potential in the modeling and treatment of various CNS diseases and disorders. However, expansion and neuronal differentiation of hNPCs in quantities necessary for these applications is not possible with current two dimensional (2-D) approaches. Here, we used a fully defined peptide substrate as the basis for a microcarrier (MC)-based suspension culture system. Several independently derived hNPC lines were cultured on MCs for multiple passages as well as efficiently differentiated to neurons. Finally, this MC-based system was used in conjunction with a low shear rotating wall vessel (RWV) bioreactor for the integrated, large-scale expansion and neuronal differentiation of hNPCs. Overall, this fully defined and scalable biomanufacturing system will facilitate the generation of hNPCs and their neuronal derivatives in quantities necessary for basic and translational applications. Statement of Significance: In this work, we developed a microcarrier (MC)-based culture system that allows for the expansion and neuronal differentiation of human pluripotent stem cell-derived neural progenitor cells (hNPCs) under defined conditions. In turn, this MC approach was implemented in a rotating wall vessel (RWV) bioreactor for the large-scale expansion and neuronal differentiation of hNPCs. This work is of significance as it overcomes current limitations of conventional two dimensional (2-D) culture systems to enable the generation of hNPCs and their neuronal derivatives in quantities required for downstream applications in disease modeling, drug screening, and regenerative medicine.
AB - Human pluripotent stem cell derived neural progenitor cells (hNPCs) have the unique properties of long-term in vitro expansion as well as differentiation into the various neurons and supporting cell types of the central nervous system (CNS). Because of these characteristics, hNPCs have tremendous potential in the modeling and treatment of various CNS diseases and disorders. However, expansion and neuronal differentiation of hNPCs in quantities necessary for these applications is not possible with current two dimensional (2-D) approaches. Here, we used a fully defined peptide substrate as the basis for a microcarrier (MC)-based suspension culture system. Several independently derived hNPC lines were cultured on MCs for multiple passages as well as efficiently differentiated to neurons. Finally, this MC-based system was used in conjunction with a low shear rotating wall vessel (RWV) bioreactor for the integrated, large-scale expansion and neuronal differentiation of hNPCs. Overall, this fully defined and scalable biomanufacturing system will facilitate the generation of hNPCs and their neuronal derivatives in quantities necessary for basic and translational applications. Statement of Significance: In this work, we developed a microcarrier (MC)-based culture system that allows for the expansion and neuronal differentiation of human pluripotent stem cell-derived neural progenitor cells (hNPCs) under defined conditions. In turn, this MC approach was implemented in a rotating wall vessel (RWV) bioreactor for the large-scale expansion and neuronal differentiation of hNPCs. This work is of significance as it overcomes current limitations of conventional two dimensional (2-D) culture systems to enable the generation of hNPCs and their neuronal derivatives in quantities required for downstream applications in disease modeling, drug screening, and regenerative medicine.
KW - Bioreactor
KW - Chemically defined peptide substrate
KW - Human neural progenitor cells
KW - Large-scale
KW - Neuronal differentiation
KW - Pluripotent stem cells
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U2 - 10.1016/j.actbio.2018.05.008
DO - 10.1016/j.actbio.2018.05.008
M3 - Article
C2 - 29775730
AN - SCOPUS:85047264896
SN - 1742-7061
VL - 74
SP - 168
EP - 179
JO - Acta Biomaterialia
JF - Acta Biomaterialia
ER -