Robotic high-throughput biomanufacturing and functional differentiation of human pluripotent stem cells.
Automation
Cell Culture Techniques
/ methods
Cell Differentiation
Cell Lineage
Cells, Cultured
Embryoid Bodies
/ cytology
Hepatocytes
/ cytology
Human Embryonic Stem Cells
/ cytology
Humans
Induced Pluripotent Stem Cells
/ cytology
Myocytes, Cardiac
/ cytology
Neurons
/ cytology
RNA-Seq
Reference Standards
Robotics
Single-Cell Analysis
Zika Virus Infection
/ pathology
CEPT cocktail
biomanufacturing
cell differentiation
high-throughput
iPS cell
mass cytometry
robotic cell culture
single-cell transcriptomics
standardization
Journal
Stem cell reports
ISSN: 2213-6711
Titre abrégé: Stem Cell Reports
Pays: United States
ID NLM: 101611300
Informations de publication
Date de publication:
14 12 2021
14 12 2021
Historique:
received:
21
08
2020
revised:
02
11
2021
accepted:
04
11
2021
pubmed:
4
12
2021
medline:
4
3
2022
entrez:
3
12
2021
Statut:
ppublish
Résumé
Efficient translation of human induced pluripotent stem cells (hiPSCs) requires scalable cell manufacturing strategies for optimal self-renewal and functional differentiation. Traditional manual cell culture is variable and labor intensive, posing challenges for high-throughput applications. Here, we established a robotic platform and automated all essential steps of hiPSC culture and differentiation under chemically defined conditions. This approach allowed rapid and standardized manufacturing of billions of hiPSCs that can be produced in parallel from up to 90 different patient- and disease-specific cell lines. Moreover, we established automated multi-lineage differentiation and generated functional neurons, cardiomyocytes, and hepatocytes. To validate our approach, we compared robotic and manual cell culture operations and performed comprehensive molecular and cellular characterizations (e.g., single-cell transcriptomics, mass cytometry, metabolism, electrophysiology) to benchmark industrial-scale cell culture operations toward building an integrated platform for efficient cell manufacturing for disease modeling, drug screening, and cell therapy.
Identifiants
pubmed: 34861164
pii: S2213-6711(21)00585-3
doi: 10.1016/j.stemcr.2021.11.004
pmc: PMC8693769
pii:
doi:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
3076-3092Commentaires et corrections
Type : UpdateOf
Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.
Références
Science. 1998 Nov 6;282(5391):1145-7
pubmed: 9804556
Nat Methods. 2006 Oct;3(10):801-6
pubmed: 16990812
Bioprocess Biosyst Eng. 2016 Dec;39(12):1847-1858
pubmed: 27503483
Nat Methods. 2011 May;8(5):424-9
pubmed: 21478862
Nat Biotechnol. 2015 Nov;33(11):1182-92
pubmed: 26501952
Stem Cell Res. 2013 Nov;11(3):1103-16
pubmed: 23973800
Nat Biotechnol. 2008 Mar;26(3):313-5
pubmed: 18278034
PLoS One. 2015 Feb 25;10(2):e0118307
pubmed: 25714340
Cytotherapy. 2019 Nov;21(11):1095-1111
pubmed: 31711733
Nat Biotechnol. 2009 Mar;27(3):275-80
pubmed: 19252484
Nat Methods. 2015 Sep;12(9):885-92
pubmed: 26237226
Cell Stem Cell. 2020 Mar 5;26(3):309-329
pubmed: 32142662
Cell Rep. 2016 Aug 9;16(6):1536-1547
pubmed: 27477285
J Biomol Screen. 2013 Mar;18(3):258-68
pubmed: 23042076
SLAS Technol. 2018 Aug;23(4):315-325
pubmed: 28574793
Stem Cell Reports. 2017 Jul 11;9(1):355-365
pubmed: 28602613
Cell Stem Cell. 2016 Oct 6;19(4):476-490
pubmed: 27618217
Nat Commun. 2015 Jun 16;6:7329
pubmed: 26076835
Cell Stem Cell. 2017 Aug 3;21(2):274-283.e5
pubmed: 28736217
J Biosci Bioeng. 2018 Jan;125(1):111-115
pubmed: 28864123
Stem Cell Reports. 2016 Mar 8;6(3):330-41
pubmed: 26923824
Nat Methods. 2021 May;18(5):528-541
pubmed: 33941937
Stem Cells. 2014 Apr;32(4):1032-42
pubmed: 24357014
Nat Commun. 2014;5:3195
pubmed: 24463987
Curr Protoc Stem Cell Biol. 2013 Sep 20;26:1G.4.1-1G.4.13
pubmed: 24510789
Nat Methods. 2020 Mar;17(3):335-342
pubmed: 32066960
Cell Stem Cell. 2015 Mar 5;16(3):323-37
pubmed: 25748935
Stem Cell Reports. 2016 Jun 14;6(6):993-1008
pubmed: 27304920
F1000Res. 2017 May 26;6:748
pubmed: 28663787
Nature. 2013 Sep 19;501(7467):373-9
pubmed: 23995685
Nat Biotechnol. 2006 Feb;24(2):185-7
pubmed: 16388305
Cell. 2018 May 17;173(5):1111-1122.e10
pubmed: 29606355
Cell Stem Cell. 2016 May 5;18(5):587-90
pubmed: 26952870
Stem Cell Reports. 2017 Apr 11;8(4):1086-1100
pubmed: 28410642
Stem Cell Reports. 2016 Sep 13;7(3):527-542
pubmed: 27569059
J Biotechnol. 2014 Mar 10;173:53-8
pubmed: 24440272
Stem Cell Reports. 2014 Dec 9;3(6):931-9
pubmed: 25496616
Cell Syst. 2019 Jul 24;9(1):35-48.e5
pubmed: 31302153
Cell. 2016 May 19;165(5):1238-1254
pubmed: 27118425
Stem Cell Reports. 2019 Apr 9;12(4):772-786
pubmed: 30827876
Biotechnol Bioeng. 2009 Apr 15;102(6):1636-44
pubmed: 19062183
Sci Rep. 2015 Nov 17;5:16647
pubmed: 26573336
Nature. 2017 May 4;545(7652):48-53
pubmed: 28445462
Stem Cell Reports. 2014 Jun 12;3(1):142-55
pubmed: 25068128
Nat Biomed Eng. 2018 Jun;2(6):362-376
pubmed: 31011198
Stem Cell Reports. 2020 Feb 11;14(2):256-270
pubmed: 31928950
Nature. 2019 May;569(7756):368-373
pubmed: 31068696
Nat Biotechnol. 2007 Jun;25(6):681-6
pubmed: 17529971
Sci Rep. 2019 Aug 23;9(1):12297
pubmed: 31444389
Nat Rev Drug Discov. 2020 Jul;19(7):463-479
pubmed: 32612263
Nat Rev Mol Cell Biol. 2013 Jun;14(6):357-68
pubmed: 23673969
Methods. 2016 May 15;101:11-20
pubmed: 26658353