Biphasic electrical field stimulation aids in tissue engineering of multicell-type cardiac organoids.
The main objectives of current work were (1) to compare the effects of monophasic or biphasic electrical field stimulation on structure and function of engineered cardiac organoids based on enriched cardiomyocytes (CM) and (2) to determine if electrical field stimulation will enhance electrical excitability of cardiac organoids based on multiple cell types. Organoids resembling cardiac myofibers were cultivated in Matrigel-coated microchannels fabricated of poly(ethylene glycol)-diacrylate. We found that field stimulation using symmetric biphasic square pulses at 2.5 V/cm, 1 Hz, 1 ms (per pulse phase) was an improved stimulation protocol, as compared to no stimulation and stimulation using monophasic square pulses of identical total amplitude and duration (5 V/cm, 1 Hz, 2 ms). This was supported by the highest success rate for synchronous contractions, low excitation threshold, the highest cell density, and the highest expression of Connexin-43 in the biphasic group. Subsequently, enriched CM were seeded on the networks of (1) cardiac fibroblasts (FB), (2) D4T endothelial cells (EC), or (3) a mixture of FB and EC that were precultured for 2 days prior to the addition of enriched CM. Biphasic field stimulation was also effective at improving electrical excitability of these cardiac organoids by improving the three-dimensional organization of the cells, increasing cellular elongation and enhancing Connexin-43 presence.
Chiu
Loraine L Y
LL
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada.
Iyer
Rohin K
RK
King
John-Paul
JP
Radisic
Milica
M
D000818:Animals; D002452:Cell Count; D002470:Cell Survival; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism; D004558:Electric Stimulation; D005786:Gene Expression Regulation; D051379:Mice; D009206:Myocardium / Q000166:cytology*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism; D009940:Organoids / Q000166:cytology*; D046509:Tissue Culture Techniques; D023822:Tissue Engineering / Q000379:methods*
D018031:Connexin 43
Pacing Clin Electrophysiol. 1990 Sep;13(9):1158-72
1700392
Circulation. 2006 Jul 4;114(1 Suppl):I72-8
16820649
Lab Chip. 2004 Oct;4(5):425-30
15472725
Endocrinology. 2004 May;145(5):2458-66
14736733
Development. 1998 Feb;125(4):725-32
9435292
Pacing Clin Electrophysiol. 1990 Oct;13(10):1268-76
1701542
Biotechnol Bioeng. 2003 May 20;82(4):403-14
12632397
Biophys J. 1992 Aug;63(2):371-86
1420884
Nat Biotechnol. 2005 Jul;23(7):879-84
15965465
Circulation. 2004 Aug 24;110(8):962-8
15302801
Heart Fail Rev. 2003 Jul;8(3):259-69
12878835
Am J Physiol. 1987 Dec;253(6 Pt 2):H1418-24
3425743
Langmuir. 2002 Apr 2;18(7):2459-62
12088033
J Biomed Mater Res A. 2009 Jun;89(3):616-31
18442120
Am Heart J. 1989 Aug;118(2):288-91
2750650
IEEE Trans Biomed Eng. 2001 Oct;48(10):1065-70
11585029
Circ Res. 2002 Nov 15;91(10):888-98
12433833
Cytobios. 1980;28(109):41-61
7428441
Tissue Eng. 2006 Aug;12(8):2077-91
16968150
J Biomed Mater Res A. 2008 Sep;86(3):713-24
18041719
Circ Res. 2002 Apr 5;90(6):671-7
11934834
Am J Physiol Heart Circ Physiol. 2004 Feb;286(2):H507-16
14551059
J Cardiovasc Electrophysiol. 2002 Apr;13(4):355-60
12033352
J Mol Cell Cardiol. 2006 Oct;41(4):633-41
16950369
Proc Natl Acad Sci U S A. 2004 Dec 28;101(52):18129-34
15604141