Persistence of intramyocardially transplanted murine induced pluripotent stem cell-derived cardiomyocytes from different developmental stages.
Cell persistence
Cell therapy
Induced pluripotent stem cell-derived cardiomyocytes
Journal
Stem cell research & therapy
ISSN: 1757-6512
Titre abrégé: Stem Cell Res Ther
Pays: England
ID NLM: 101527581
Informations de publication
Date de publication:
08 01 2021
08 01 2021
Historique:
received:
20
08
2020
accepted:
09
12
2020
entrez:
9
1
2021
pubmed:
10
1
2021
medline:
6
7
2021
Statut:
epublish
Résumé
Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) are regarded as promising cell type for cardiac cell replacement therapy, but it is not known whether the developmental stage influences their persistence and functional integration in the host tissue, which are crucial for a long-term therapeutic benefit. To investigate this, we first tested the cell adhesion capability of murine iPSC-CM in vitro at three different time points during the differentiation process and then examined cell persistence and quality of electrical integration in the infarcted myocardium in vivo. To test cell adhesion capabilities in vitro, iPSC-CM were seeded on fibronectin-coated cell culture dishes and decellularized ventricular extracellular matrix (ECM) scaffolds. After fixed periods of time, stably attached cells were quantified. For in vivo experiments, murine iPSC-CM expressing enhanced green fluorescent protein was injected into infarcted hearts of adult mice. After 6-7 days, viable ventricular tissue slices were prepared to enable action potential (AP) recordings in transplanted iPSC-CM and surrounding host cardiomyocytes. Afterwards, slices were lysed, and genomic DNA was prepared, which was then used for quantitative real-time PCR to evaluate grafted iPSC-CM count. The in vitro results indicated differences in cell adhesion capabilities between day 14, day 16, and day 18 iPSC-CM with day 14 iPSC-CM showing the largest number of attached cells on ECM scaffolds. After intramyocardial injection, day 14 iPSC-CM showed a significant higher cell count compared to day 16 iPSC-CM. AP measurements revealed no significant difference in the quality of electrical integration and only minor differences in AP properties between d14 and d16 iPSC-CM. The results of the present study demonstrate that the developmental stage at the time of transplantation is crucial for the persistence of transplanted iPSC-CM. iPSC-CM at day 14 of differentiation showed the highest persistence after transplantation in vivo, which may be explained by a higher capability to adhere to the extracellular matrix.
Sections du résumé
BACKGROUND
Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) are regarded as promising cell type for cardiac cell replacement therapy, but it is not known whether the developmental stage influences their persistence and functional integration in the host tissue, which are crucial for a long-term therapeutic benefit. To investigate this, we first tested the cell adhesion capability of murine iPSC-CM in vitro at three different time points during the differentiation process and then examined cell persistence and quality of electrical integration in the infarcted myocardium in vivo.
METHODS
To test cell adhesion capabilities in vitro, iPSC-CM were seeded on fibronectin-coated cell culture dishes and decellularized ventricular extracellular matrix (ECM) scaffolds. After fixed periods of time, stably attached cells were quantified. For in vivo experiments, murine iPSC-CM expressing enhanced green fluorescent protein was injected into infarcted hearts of adult mice. After 6-7 days, viable ventricular tissue slices were prepared to enable action potential (AP) recordings in transplanted iPSC-CM and surrounding host cardiomyocytes. Afterwards, slices were lysed, and genomic DNA was prepared, which was then used for quantitative real-time PCR to evaluate grafted iPSC-CM count.
RESULTS
The in vitro results indicated differences in cell adhesion capabilities between day 14, day 16, and day 18 iPSC-CM with day 14 iPSC-CM showing the largest number of attached cells on ECM scaffolds. After intramyocardial injection, day 14 iPSC-CM showed a significant higher cell count compared to day 16 iPSC-CM. AP measurements revealed no significant difference in the quality of electrical integration and only minor differences in AP properties between d14 and d16 iPSC-CM.
CONCLUSION
The results of the present study demonstrate that the developmental stage at the time of transplantation is crucial for the persistence of transplanted iPSC-CM. iPSC-CM at day 14 of differentiation showed the highest persistence after transplantation in vivo, which may be explained by a higher capability to adhere to the extracellular matrix.
Identifiants
pubmed: 33419458
doi: 10.1186/s13287-020-02089-5
pii: 10.1186/s13287-020-02089-5
pmc: PMC7792075
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
46Références
Biomaterials. 2014 Oct;35(30):8528-39
pubmed: 25043570
Ann Thorac Surg. 2005 Nov;80(5):1779-86
pubmed: 16242455
J Pharmacol Exp Ther. 2019 Sep;370(3):761-771
pubmed: 30728248
Cell Physiol Biochem. 2007;20(6):837-46
pubmed: 17982265
Cell Stem Cell. 2014 Dec 4;15(6):750-61
pubmed: 25479750
J Cell Biol. 1994 Feb;124(4):619-26
pubmed: 8106557
J Nucl Med. 2007 Oct;48(10):1708-14
pubmed: 17909258
Circ Res. 2007 Aug 31;101(5):484-92
pubmed: 17641227
Circulation. 2002 May 21;105(20):2435-41
pubmed: 12021233
J Neurobiol. 1998 Oct;37(1):37-59
pubmed: 9777731
Cell. 2007 Nov 30;131(5):861-72
pubmed: 18035408
Transplantation. 2019 Aug;103(8):1582-1590
pubmed: 31107828
Cell Physiol Biochem. 2019;52(6):1309-1324
pubmed: 31050280
Stem Cells Dev. 2010 Oct;19(10):1589-99
pubmed: 20175666
Acta Biomater. 2017 Mar 1;50:207-219
pubmed: 27993639
Nature. 2007 Jul 19;448(7151):313-7
pubmed: 17554338
BMC Bioinformatics. 2017 Nov 29;18(1):529
pubmed: 29187165
Circulation. 2008 Jul 29;118(5):498-506
pubmed: 18625891
Circulation. 2004 Sep 14;110(11 Suppl 1):II219-24
pubmed: 15364866
Cell Physiol Biochem. 2015;35(1):305-14
pubmed: 25591772
Cell Stem Cell. 2013 Jan 3;12(1):127-37
pubmed: 23168164
J Vis Exp. 2017 Jun 4;(124):
pubmed: 28605368
Mol Pharm. 2011 Oct 3;8(5):1573-81
pubmed: 21542647
Biomaterials. 2015 Dec;73:1-11
pubmed: 26378976
Nat Biotechnol. 2007 Oct;25(10):1177-81
pubmed: 17724450
Cell Stem Cell. 2011 Feb 4;8(2):228-40
pubmed: 21295278
Int J Cardiol. 2014 Feb 15;171(3):e122-4
pubmed: 24439862
Cell Physiol Biochem. 2006;18(1-3):1-8
pubmed: 16914885
J Mol Cell Cardiol. 2002 Feb;34(2):107-16
pubmed: 11851351
Eur J Nucl Med Mol Imaging. 2013 Oct;40(11):1730-8
pubmed: 23860738
Nature. 2018 May;557(7707):619-620
pubmed: 29844563
Eur Heart J. 2017 Jan 14;38(3):184-186
pubmed: 28158468
Biomaterials. 2014 Aug;35(26):7374-85
pubmed: 24889032
PLoS One. 2012;7(8):e44228
pubmed: 22952932
Circ Cardiovasc Imaging. 2016 Nov;9(11):
pubmed: 27903535
Cell Physiol Biochem. 2009;24(1-2):73-86
pubmed: 19590195
Sci Transl Med. 2016 Nov 2;8(363):363ra148
pubmed: 27807283
Stem Cell Res. 2016 Sep;17(2):266-272
pubmed: 27879210
Circ Res. 2009 Feb 27;104(4):e30-41
pubmed: 19213953
Artif Organs. 2010 Mar;34(3):206-10
pubmed: 20447045
J Am Coll Cardiol. 2003 Dec 17;42(12):2063-9
pubmed: 14680727
Mol Pharm. 2013 Sep 3;10(9):3425-32
pubmed: 23879836
Eur Heart J. 2006 May;27(9):1114-22
pubmed: 16510464
PLoS One. 2012;7(9):e45963
pubmed: 23029342
Circulation. 2013 Feb 12;127(6):749-56
pubmed: 23401116
Stem Cells Dev. 2016 Oct 1;25(19):1397-406
pubmed: 27484788
Circ Res. 1996 Jul;79(1):79-85
pubmed: 8925572
Circ Res. 1997 Jul;81(1):120-7
pubmed: 9201035
Cardiovasc Res. 2013 Dec 1;100(3):432-40
pubmed: 24042016
Cell Transplant. 2017 Jan 24;26(1):157-170
pubmed: 27539827
Circulation. 2005 Aug 30;112(9 Suppl):I150-6
pubmed: 16159808
Circulation. 1999 Jul 13;100(2):193-202
pubmed: 10402450
Circ Res. 2014 Jan 31;114(3):572-586
pubmed: 24481847
Biomaterials. 2014 Jan;35(1):133-42
pubmed: 24099706