Stem cell function is conserved during short-term storage of cultured epidermal cell sheets at 12°C.
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2020
2020
Historique:
received:
30
10
2019
accepted:
10
04
2020
entrez:
21
5
2020
pubmed:
21
5
2020
medline:
30
7
2020
Statut:
epublish
Résumé
Transplantation of cultured epidermal cell sheets (CES) can be life-saving for patients with large area burns. CES have also been successfully used to regenerate eye and urethral epithelia in animal models. Short-term storage aims to extend the transplantation window, offers flexibility in timing surgery and allows testing of CES quality, phenotype and sterility. This study investigated extended CES storage and explored the effect of additional re-incubation recovery time following storage. The proliferative quality of stored confluent versus pre-confluent CES was also investigated using functional testing. CES were stored at 12°C and results compared to non-stored control CES. Investigation of timepoints during 15 days storage revealed that viability began to deteriorate by day 11 and was associated with increased lactate in the storage medium. The percentage of apoptotic cells also significantly increased by day 11. Flow cytometry analysis of integrin β1 expression and cell size indicated best retention of stem cells at 7 days of storage. Functional testing of pre-confluent and confluent cells following 7 days storage showed that pre-confluent cells responded well to 1-day re-incubation after storage; they became highly prolific, increasing in number by ~67%. Conversely, proliferation in stored confluent cells declined by ~50% with 1-day re-incubation. Pre-confluent stored CES also had far superior stem cell colony forming efficiency (CFE) performance compared to the confluent group. Re-incubation improved CFE in both groups, but the pre-confluent group again out-performed the confluent group with significantly more colonies. In conclusion, a maximum storage period of 7 days is recommended. Use of pre-confluent cells and one day recovery incubation greatly improves viability, colony-forming ability and proliferation of cells stored for 7 days at 12°C. Thus, these recommendations should be considered under culture and storage of high-quality CES for clinical use.
Identifiants
pubmed: 32433698
doi: 10.1371/journal.pone.0232270
pii: PONE-D-19-30258
pmc: PMC7239464
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0232270Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
Références
Cell. 1993 May 21;73(4):713-24
pubmed: 8500165
PLoS One. 2014 Aug 29;9(8):e105808
pubmed: 25170754
Semin Cell Dev Biol. 2012 Oct;23(8):937-44
pubmed: 23036530
Mol Reprod Dev. 2007 Mar;74(3):386-96
pubmed: 16998851
PLoS One. 2013 Jul 23;8(7):e69687
pubmed: 23936078
Cell. 1975 Nov;6(3):331-43
pubmed: 1052771
J Burn Care Rehabil. 1992 Jan-Feb;13(1):130-7
pubmed: 1572844
J Vis Exp. 2011 Mar 13;(49):
pubmed: 21445039
Cryobiology. 1999 Sep;39(2):158-68
pubmed: 10529309
Curr Eye Res. 2016 Jun;41(6):757-68
pubmed: 26398483
Proc Natl Acad Sci U S A. 1985 Aug;82(16):5390-4
pubmed: 2410922
Proc Natl Acad Sci U S A. 1987 Apr;84(8):2302-6
pubmed: 2436229
J Invest Dermatol. 1995 Feb;104(2):271-6
pubmed: 7530273
Acta Naturae. 2015 Jan-Mar;7(1):70-7
pubmed: 25927003
Regen Med. 2006 Jan;1(1):45-57
pubmed: 17465819
N Engl J Med. 1984 Aug 16;311(7):448-51
pubmed: 6379456
Burns. 1998 Nov;24(7):591-3
pubmed: 9882054
Sci Rep. 2017 Aug 15;7(1):8206
pubmed: 28811665
Stem Cells. 2011 Aug;29(8):1256-68
pubmed: 21674699
Int Wound J. 2013 Feb;10(1):6-12
pubmed: 22958654
J Cell Physiol. 2010 Oct;225(1):15-9
pubmed: 20506173