A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application.
Clinical application
DMEM with antibiotics, (DMEM-AB)
Dulbecco's modified Eagle's medium, (DMEM)
Epithelial cell sheet
Minimum trypsinization
Primary cell culture
epithelial growth factor, (EGF)
keratinocyte culture medium, (KCM)
poly(N-isopropyl acrylamide), (PIPAAm)
polymerase chain reaction, (PCR)
Journal
Regenerative therapy
ISSN: 2352-3204
Titre abrégé: Regen Ther
Pays: Netherlands
ID NLM: 101709085
Informations de publication
Date de publication:
Jun 2020
Jun 2020
Historique:
received:
29
07
2019
revised:
08
11
2019
accepted:
30
11
2019
entrez:
29
1
2020
pubmed:
29
1
2020
medline:
29
1
2020
Statut:
epublish
Résumé
Cultured stratified epithelial cell sheets have been clinically utilized as transplantable grafts for the regeneration of epithelial tissues, such as the esophagus, cornea, skin, and intraoral cavity. These cell sheets are expected to gain widespread use as regenerative medicine products and save many patients. For this purpose, establishing and disseminating the stale protocol of fabricating the cell sheet is crucial. The fabrication of cultured stratified epithelial cell sheets consists of many important steps, and since the patients' epithelial cell conditions vary widely and are sometimes unstable, the qualities of the epithelial cell grafts are likewise potentially unstable. Therefore, in this paper, we report the stable protocol for fabrication of the transplantable cell sheet particularly from patient-derived oral mucosal tissues. Serum extracted from blood and buccal mucosal tissue were collected in Nagasaki University and transported to Tokyo Women's Medical University. Oral mucosal epithelial cells were collected by minimum trypsin method, and this treatment was studied whether to be a critical procedure. After 14 days cultivation, cultured cells were examined whether to be transplantable as cell sheets. We successfully transported buccal mucosal tissue and serum without damage and contamination. Oral mucosal epithelial cells were collected with high viability by minimum trypsin method. Finally, we succeeded to stably fabricate oral mucosal epithelial cell sheets in all 10 patients. We established a stable protocol for the fabrication of human oral mucosal epithelial cell sheets and their transportation in clinical settings in this study. These methodologies could also be basis for transplantation therapy using cultured cell sheets of various types other than oral mucosal epithelial cell and will contribute largely to the future development of regenerative medicine.
Identifiants
pubmed: 31988998
doi: 10.1016/j.reth.2019.11.007
pii: S2352-3204(19)30157-9
pmc: PMC6970131
doi:
Types de publication
Journal Article
Langues
eng
Pagination
87-94Informations de copyright
© 2020 The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V.
Déclaration de conflit d'intérêts
Yamato Masayuki is a stakeholder of CellSeed Inc., and Tokyo Women's Medical University receives research funding from CellSeed Inc. Yamato Masayuki is also an advisor of Helios and Nippi. Teruo Okano is a founder of CellSeed Inc. which has licenses for certain cell sheet-related technologies and patents from Tokyo Women's Medical University.
Références
Biomaterials. 1995 Mar;16(4):297-303
pubmed: 7772669
Methods Cell Biol. 1980;21A:229-54
pubmed: 6157968
Gastroenterology. 2012 Sep;143(3):582-588.e2
pubmed: 22561054
N Engl J Med. 2004 Sep 16;351(12):1187-96
pubmed: 15371576
Br J Ophthalmol. 2004 Oct;88(10):1280-4
pubmed: 15377551
Exp Cell Res. 1974 Mar 15;84(1):207-18
pubmed: 4361714
Br J Dermatol. 1983 May;108(5):555-60
pubmed: 6342649
Regen Ther. 2018 Dec 17;10:36-45
pubmed: 30581895
In Vitro. 1974 Jul-Aug;10:97-111
pubmed: 4143484
Nature. 1977 Feb 3;265(5593):421-4
pubmed: 299924
Cell. 1978 Nov;15(3):801-11
pubmed: 83196
Hum Cell. 1996 Mar;9(1):91-6
pubmed: 9183636
Head Neck. 2000 Oct;22(7):658-61
pubmed: 11002319
Int J Cancer. 1969 Mar 15;4(2):159-65
pubmed: 4186775
J Artif Organs. 2006;9(3):185-91
pubmed: 16998704
Regen Ther. 2015 Feb 18;1:1-4
pubmed: 31245435
J Biomed Mater Res A. 2012 Oct;100(10):2849-54
pubmed: 22623227
Am J Ophthalmol. 2011 Nov;152(5):739-47.e1
pubmed: 21803325
Int J Oral Maxillofac Surg. 2003 Apr;32(2):188-97
pubmed: 12729781
Gastrointest Endosc. 2012 Oct;76(4):873-81
pubmed: 22867446
Sci Rep. 2017 Dec 12;7(1):17460
pubmed: 29234120
Br J Dermatol. 2003 Aug;149(2):255-60
pubmed: 12932229
Transplantation. 1990 Sep;50(3):454-9
pubmed: 2402795
Invest Ophthalmol Vis Sci. 2012 Mar 13;53(3):1325-31
pubmed: 22064987
Proc Natl Acad Sci U S A. 1979 Nov;76(11):5665-8
pubmed: 293669
J Biomed Mater Res. 1993 Oct;27(10):1243-51
pubmed: 8245039