Functional human iPSC-derived alveolar-like cells cultured in a miniaturized 96‑Transwell air-liquid interface model.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
23 08 2021
23 08 2021
Historique:
received:
20
05
2021
accepted:
11
08
2021
entrez:
24
8
2021
pubmed:
25
8
2021
medline:
5
11
2021
Statut:
epublish
Résumé
In order to circumvent the limited access and donor variability of human primary alveolar cells, directed differentiation of human pluripotent stem cells (hiPSCs) into alveolar-like cells, provides a promising tool for respiratory disease modeling and drug discovery assays. In this work, a unique, miniaturized 96-Transwell microplate system is described where hiPSC-derived alveolar-like cells were cultured at an air-liquid interface (ALI). To this end, hiPSCs were differentiated into lung epithelial progenitor cells (LPCs) and subsequently matured into a functional alveolar type 2 (AT2)-like epithelium with monolayer-like morphology. AT2-like cells cultured at the physiological ALI conditions displayed characteristics of AT2 cells with classical alveolar surfactant protein expressions and lamellar-body like structures. The integrity of the epithelial barriers between the AT2-like cells was confirmed by applying a custom-made device for 96-parallelized transepithelial electric resistance (TEER) measurements. In order to generate an IPF disease-like phenotype in vitro, the functional AT2-like cells were stimulated with cytokines and growth factors present in the alveolar tissue of IPF patients. The cytokines stimulated the secretion of pro-fibrotic biomarker proteins both on the mRNA (messenger ribonucleic acid) and protein level. Thus, the hiPSC-derived and cellular model system enables the recapitulation of certain IPF hallmarks, while paving the route towards a miniaturized medium throughput approach of pharmaceutical drug discovery.
Identifiants
pubmed: 34426605
doi: 10.1038/s41598-021-96565-4
pii: 10.1038/s41598-021-96565-4
pmc: PMC8382767
doi:
Substances chimiques
Biomarkers
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
17028Informations de copyright
© 2021. The Author(s).
Références
Cell Stem Cell. 2017 Oct 5;21(4):472-488.e10
pubmed: 28965766
Am J Respir Cell Mol Biol. 2014 Apr;50(4):825-37
pubmed: 24325577
Thorax. 2015 Jan;70(1):48-56
pubmed: 25217476
Methods Mol Med. 2001;56:137-46
pubmed: 21336897
Am Rev Respir Dis. 1989 Dec;140(6):1693-703
pubmed: 2604297
FASEB J. 2020 Jun;34(6):7825-7846
pubmed: 32297676
Respir Med. 2017 Oct;131:49-57
pubmed: 28947042
Toxicol Lett. 2013 May 10;219(1):49-58
pubmed: 23470867
Cell Cycle. 2010 Jul 15;9(14):2769-76
pubmed: 20676040
Respir Res. 2001;2(1):33-46
pubmed: 11686863
Pharmacol Ther. 2017 Dec;180:90-98
pubmed: 28642115
Am Rev Respir Dis. 1983 Aug;128(2 Pt 2):S42-6
pubmed: 6881707
Am J Physiol Lung Cell Mol Physiol. 2015 Jun 15;308(12):L1212-23
pubmed: 25888574
SLAS Discov. 2017 Mar;22(3):274-286
pubmed: 28231034
Am J Physiol. 1994 Mar;266(3 Pt 1):L278-86
pubmed: 8166297
Eur Respir Rev. 2019 Feb 27;28(151):
pubmed: 30814139
Cell. 2021 Feb 4;184(3):845-846
pubmed: 33545039
Proc Am Thorac Soc. 2008 Aug 15;5(6):717-22
pubmed: 18684724
Clin Exp Immunol. 2011 Dec;166(3):346-51
pubmed: 22059992
Cell. 2006 Aug 25;126(4):663-76
pubmed: 16904174
J Clin Invest. 2015 Feb;125(2):521-38
pubmed: 25562319
J Invest Dermatol. 2002 Feb;118(2):211-5
pubmed: 11841535
Neuron. 2018 Nov 21;100(4):783-797
pubmed: 30465765
Respirology. 2012 Jul;17(5):814-20
pubmed: 22515655
Proc Am Thorac Soc. 2006 Jun;3(4):357-63
pubmed: 16738201
Eur Respir J. 2010 Apr;35(4):894-903
pubmed: 19741028
Differentiation. 2018 May - Jun;101:16-24
pubmed: 29626713
Development. 2015 Jan 1;142(1):13-6
pubmed: 25516965
Biosci Rep. 2017 Nov 21;37(6):
pubmed: 29026006
PLoS One. 2010 Aug 13;5(8):e12148
pubmed: 20730054
Drug Discov Today. 2019 Feb;24(2):526-532
pubmed: 30359770
Cell Stem Cell. 2020 Dec 3;27(6):962-973.e7
pubmed: 32979316
Methods Mol Biol. 2019;1994:101-115
pubmed: 31124108
J Pathol. 2008 Jan;214(2):199-210
pubmed: 18161745
J Clin Invest. 2012 Aug;122(8):2724-30
pubmed: 22850882
J Histochem Cytochem. 2007 Jan;55(1):71-83
pubmed: 16982851
Nat Protoc. 2015 Mar;10(3):413-25
pubmed: 25654758
Nat Cell Biol. 2017 May;19(5):542-549
pubmed: 28436965
Annu Rev Physiol. 2003;65:669-95
pubmed: 12428023
Sci Rep. 2019 Jan 24;9(1):505
pubmed: 30679658
Biochem Biophys Res Commun. 2007 Jun 1;357(2):402-7
pubmed: 17420005
Tissue Eng Part A. 2015 Feb;21(3-4):669-82
pubmed: 25316003
Biotechnol Prog. 2010 May-Jun;26(3):827-37
pubmed: 20077485
PLoS One. 2012;7(3):e33165
pubmed: 22470441
Exp Cell Res. 1998 Sep 15;243(2):359-66
pubmed: 9743595
N Engl J Med. 2018 May 10;378(19):1811-1823
pubmed: 29742380
Cell Stem Cell. 2017 Jun 1;20(6):844-857.e6
pubmed: 28366587
Clin Respir J. 2016 Mar;10(2):133-41
pubmed: 25047066
Stem Cell Reports. 2014 Sep 9;3(3):394-403
pubmed: 25241738
Am J Respir Cell Mol Biol. 2018 May;58(5):547-548
pubmed: 29714627
J Cell Mol Med. 2016 Nov;20(11):2064-2077
pubmed: 27306439
Sci Rep. 2020 Mar 26;10(1):5499
pubmed: 32218519
Nat Biotechnol. 2005 Dec;23(12):1534-41
pubmed: 16258519
Nat Methods. 2017 Nov;14(11):1097-1106
pubmed: 28967890
Biotechnol Prog. 2010 May-Jun;26(3):781-8
pubmed: 20014103
Dev Cell. 2010 Jan 19;18(1):8-23
pubmed: 20152174
Nature. 2018 Mar 8;555(7695):251-255
pubmed: 29489752
J Clin Invest. 2013 Nov;123(11):4950-62
pubmed: 24135142
Nat Biotechnol. 2012 Sep;30(9):876-82
pubmed: 22922672
Nat Protoc. 2014;9(6):1514-31
pubmed: 24874816
Nat Protoc. 2019 Dec;14(12):3303-3332
pubmed: 31732721
Respir Res. 2002;3:3
pubmed: 11806838
PLoS One. 2016 Oct 28;11(10):e0164438
pubmed: 27792742
Nat Rev Mol Cell Biol. 2020 Oct;21(10):571-584
pubmed: 32636524
Autoimmun Rev. 2014 Oct;13(10):1020-5
pubmed: 25182202
Nat Biotechnol. 2011 Mar;29(3):267-72
pubmed: 21358635
Science. 2014 Dec 19;346(6216):1452-3
pubmed: 25525228
J Clin Invest. 2017 Oct 2;127(10):3675-3688
pubmed: 28872461
J Biol Chem. 2002 Jun 28;277(26):23573-81
pubmed: 11950839
Int J Cancer. 1976 Jan 15;17(1):62-70
pubmed: 175022
Annu Rev Pathol. 2015;10:371-93
pubmed: 25621661
Clin Rev Allergy Immunol. 2011 Apr;40(2):117-34
pubmed: 20838937
Int J Mol Sci. 2020 Mar 25;21(7):
pubmed: 32218238
Am J Respir Cell Mol Biol. 2017 Apr;56(4):432-442
pubmed: 27854509
Biochim Biophys Acta. 1981 Dec 23;666(3):382-93
pubmed: 6895699
Front Med (Lausanne). 2019 Nov 15;6:265
pubmed: 31803747
J Dermatol Sci. 2004 Aug;35(2):83-92
pubmed: 15265520
Curr Protoc Stem Cell Biol. 2020 Mar;52(1):e108
pubmed: 32159928
J Interferon Cytokine Res. 2009 Jun;29(6):313-26
pubmed: 19441883
Stem Cell Reports. 2015 Apr 14;4(4):569-77
pubmed: 25772471
J Immunol. 2000 Aug 15;165(4):2214-21
pubmed: 10925309
Am J Respir Cell Mol Biol. 2013 Aug;49(2):167-79
pubmed: 23526222
Sci Rep. 2020 Aug 3;10(1):13022
pubmed: 32747751
bioRxiv. 2020 Aug 06;:
pubmed: 32637964
Nat Protoc. 2015 Mar;10(3):363-81
pubmed: 25654755
Eur J Pharmacol. 2006 Mar 8;533(1-3):133-44
pubmed: 16487964
Tissue Eng Part A. 2009 Nov;15(11):3351-65
pubmed: 19388834
Mol Pharmacol. 2002 Jul;62(1):65-74
pubmed: 12065756
Front Med (Lausanne). 2018 Feb 05;5:10
pubmed: 29459894
Stem Cell Res Ther. 2021 Feb 25;12(1):151
pubmed: 33632328
Mol Cell Proteomics. 2009 Mar;8(3):443-50
pubmed: 18952599
Comp Biochem Physiol A Mol Integr Physiol. 2001 May;129(1):245-60
pubmed: 11369549
Cell Stem Cell. 2020 Apr 2;26(4):593-608.e8
pubmed: 32004478
Bioessays. 2015 Sep;37(9):1028-37
pubmed: 26201286
Respiration. 2013;86(6):441-52
pubmed: 24356558
Lab Invest. 2011 Mar;91(3):363-78
pubmed: 21079581
Development. 2014 Feb;141(3):502-13
pubmed: 24449833
Stem Cells Transl Med. 2014 Jan;3(1):7-17
pubmed: 24300555
J Clin Invest. 2013 Jul;123(7):3025-36
pubmed: 23921127
Elife. 2015 Mar 24;4:
pubmed: 25803487
Proc Natl Acad Sci U S A. 2014 Apr 29;111(17):E1723-30
pubmed: 24706852
Respir Res. 2015 Sep 29;16:120
pubmed: 26415518
Development. 2018 Aug 15;145(16):
pubmed: 30111617
Curr Transplant Rep. 2015 Mar 1;2(1):81-89
pubmed: 26052480
Adv Exp Med Biol. 2018;1071:109-114
pubmed: 30357741
Molecules. 2018 Jun 05;23(6):
pubmed: 29874809