Comparison of promoter, DNA vector, and cationic carrier for efficient transfection of hMSCs from multiple donors and tissue sources.
DNA vector
gene delivery
lipofection
nonviral
polymer-mediated gene delivery
screen
stem cells
transfection
Journal
Molecular therapy. Nucleic acids
ISSN: 2162-2531
Titre abrégé: Mol Ther Nucleic Acids
Pays: United States
ID NLM: 101581621
Informations de publication
Date de publication:
03 Dec 2021
03 Dec 2021
Historique:
received:
13
01
2021
accepted:
25
06
2021
entrez:
13
9
2021
pubmed:
14
9
2021
medline:
14
9
2021
Statut:
epublish
Résumé
Human mesenchymal stem cells (hMSCs) are primary cells with high clinical relevance that could be enhanced through genetic modification. However, gene delivery, particularly through nonviral routes, is inefficient. To address the shortcomings of nonviral gene delivery to hMSCs, our lab has previously demonstrated that pharmacological "priming" of hMSCs with clinically approved drugs can increase transfection in hMSCs by modulating transfection-induced cytotoxicity. However, even with priming, hMSC transfection remains inefficient for clinical applications. This work takes a complementary approach to addressing the challenges of transfecting hMSCs by systematically investigating key transfection parameters for their effect on transgene expression. Specifically, we investigated two promoters (cytomegalovirus [CMV] and elongation factor 1 alpha), four DNA vectors (plasmid, plasmid with no F1 origin, minicircle, and mini-intronic plasmid), two cationic carriers (Lipofectamine 3000 and Turbofect), and four donors of hMSCs from two tissues (adipose and bone marrow) for efficient hMSC transfection. Following systematic comparison of each variable, we identified adipose-derived hMSCs transfected with mini-intronic plasmids containing the CMV promoter delivered using Lipofectamine 3000 as the parameters that produced the highest transfection levels. The data presented in this work can guide the development of other hMSC transfection systems with the goal of producing clinically relevant, genetically modified hMSCs.
Identifiants
pubmed: 34513295
doi: 10.1016/j.omtn.2021.06.018
pii: S2162-2531(21)00159-1
pmc: PMC8413668
doi:
Types de publication
Journal Article
Langues
eng
Pagination
81-93Subventions
Organisme : NIBIB NIH HHS
ID : DP2 EB025760
Pays : United States
Informations de copyright
© 2021 The Author(s).
Déclaration de conflit d'intérêts
The authors declare no competing interests.
Références
Nat Protoc. 2011 Jan;6(1):78-88
pubmed: 21212777
Gene Ther. 2004 May;11(10):856-64
pubmed: 15029228
Genome Biol. 2018 Sep 14;19(1):133
pubmed: 30217220
Cancer Gene Ther. 2019 Jul;26(7-8):183-194
pubmed: 30100607
Mol Ther. 2012 Nov;20(11):2111-9
pubmed: 22565847
Mol Ther. 2005 Sep;12(3):528-36
pubmed: 16099414
Mol Ther. 2013 May;21(5):954-63
pubmed: 23459514
Biomaterials. 2016 Sep;101:310-20
pubmed: 27315214
Int J Mol Sci. 2020 Aug 24;21(17):
pubmed: 32847094
Mol Ther Methods Clin Dev. 2014 Sep;2014(1):
pubmed: 25279386
J Cell Biochem. 2018 Nov;119(11):8723-8736
pubmed: 30074262
Biotechnol Rep (Amst). 2016 Jun 16;11:53-61
pubmed: 28352540
Mol Ther. 2016 Feb;24(2):331-341
pubmed: 26478250
Biosci Rep. 2015 Apr 28;35(2):
pubmed: 25797907
J Mol Med (Berl). 2011 May;89(5):515-29
pubmed: 21301798
Gene Ther. 2009 Apr;16(4):533-46
pubmed: 19129861
Genes Dev. 2002 Nov 1;16(21):2792-9
pubmed: 12414732
Aging Med (Milton). 2019 Sep;2(3):142-146
pubmed: 31667462
Bratisl Lek Listy. 2018;119(11):701-705
pubmed: 30686003
Mol Ther. 2003 Sep;8(3):495-500
pubmed: 12946323
Periodontol 2000. 2013 Oct;63(1):198-216
pubmed: 23931061
Nat Genet. 2004 Sep;36(9):1014-8
pubmed: 15314641
Exp Hematol. 2008 Oct;36(10):1354-1369
pubmed: 18657893
CellR4 Repair Replace Regen Reprogram. 2017;5(5):
pubmed: 30505879
Theranostics. 2014 Jan 15;4(3):240-55
pubmed: 24505233
Genes Dev. 1990 Sep;4(9):1552-9
pubmed: 1701407
J Biol Eng. 2019 Jan 18;13:7
pubmed: 30675180
Genes (Basel). 2017 Feb 10;8(2):
pubmed: 28208635
J Biol Eng. 2020 May 19;14:16
pubmed: 32467728
J Virol. 1989 Aug;63(8):3284-95
pubmed: 2746731
Curr Gene Ther. 2011 Feb;11(1):46-57
pubmed: 21182464
Nat Biotechnol. 2010 Dec;28(12):1287-9
pubmed: 21102455
Aging Dis. 2020 Mar 9;11(2):216-228
pubmed: 32257537
Stem Cells Int. 2018 Dec 24;2018:1310904
pubmed: 30675166
Biotechnol Bioeng. 2019 Feb;116(2):427-443
pubmed: 30450542
Stem Cell Res Ther. 2018 Jun 19;9(1):168
pubmed: 29921311
Adv Biomed Res. 2012;1:27
pubmed: 23210086
Stem Cell Res Ther. 2016 Apr 01;7:48
pubmed: 27036881
EMBO J. 1998 Apr 1;17(7):2107-21
pubmed: 9524132
Curr Gene Ther. 2020;20(1):55-63
pubmed: 32338217
Chem Biol. 2010 Jun 25;17(6):549-50
pubmed: 20609403
J Cell Mol Med. 2011 Sep;15(9):1989-98
pubmed: 20629995
Gene Ther. 1997 Dec;4(12):1341-9
pubmed: 9472558
EMBO J. 2001 Sep 3;20(17):4987-97
pubmed: 11532962
J Cell Physiol. 2018 May;233(5):3982-3999
pubmed: 28926091
Hum Gene Ther. 1999 Sep 1;10(13):2153-61
pubmed: 10498247
Int J Mol Sci. 2017 Apr 12;18(4):
pubmed: 28417917
RNA. 2003 May;9(5):607-17
pubmed: 12702819
Mol Ther Methods Clin Dev. 2020 Jul 23;18:713-722
pubmed: 32913879
Cytotherapy. 2018 Mar;20(3):343-360
pubmed: 29396254
Trends Mol Med. 2012 Feb;18(2):128-34
pubmed: 22118960
Stem Cell Rev Rep. 2017 Dec;13(6):725-740
pubmed: 28815481
Nat Methods. 2010 Mar;7(3):197-9
pubmed: 20139967
Sci Rep. 2015 Jan 28;5:8081
pubmed: 25628230
Oncoimmunology. 2016 Sep 2;5(10):e1223002
pubmed: 27853647
Vaccine. 2009 Oct 30;27(46):6454-9
pubmed: 19559109
Exp Biol Med (Maywood). 2020 Apr;245(7):606-619
pubmed: 32183552
Nature. 2016 Dec 1;540(7631):144-149
pubmed: 27851729