Journal article
Human Epithelial Stem Cells Persist Within Tissue-Engineered Skin Produced by the Self-Assembly Approach
Tissue engineering. Part A, Vol.19(7-8), pp.1023-1038
04/2013
DOI: 10.1089/ten.tea.2012.0117
PMID: 23173810
Abstract
To adequately and permanently restore organ function after grafting, human tissue-engineered skin substitutes (TESs) must ultimately contain and preserve functional epithelial stem cells (SCs). It is therefore essential that a maximum of SCs be preserved during each in vitro step leading to the production of TESs such as the culture process and the elaboration of a skin cell bank by cryopreservation. To investigate the presence and functionality of epithelial SCs within the human TESs made by the self-assembly approach, slow-cycling cells were identified using 5'-bromo-2'-deoxyuridine (BrdU) in the three-dimensional construct. A subset of basal epithelial cells retained the BrdU label and was positive for the SC-associated marker keratin 19 within TESs after a chase of 21 days in culture post-BrdU labeling. Moreover, keratinocytes harvested from TESs gave rise to SC-like colonies in secondary monolayer subcultures, indicating that SCs were preserved within TESs. To evaluate the effect of cryopreservation with dimethyl sulfoxide and storage in liquid nitrogen on SCs, human epithelial cells were extracted from skin samples, amplified in culture, and used to produce TESs, before cryopreservation as well as after thawing. We found that the proportion and the growth potential of epithelial SCs in monolayer culture and in TESs remained constant before and after cryopreservation. Further, the functionality of these substitutes was demonstrated by successfully grafting human TESs on athymic mice for 6 months. We conclude that human epithelial skin SCs are adequately preserved upon human tissue reconstruction. Thus, these TESs produced by the self-assembly approach are suitable for clinical applications.
Details
- Title: Subtitle
- Human Epithelial Stem Cells Persist Within Tissue-Engineered Skin Produced by the Self-Assembly Approach
- Creators
- Amélie Lavoie - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaClaudia Fugère - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaAnnie Beauparlant - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaBenjamin Goyer - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaDanielle Larouche - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaClaudie Paquet - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaMaxime Desgagné - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaSarah Sauvé - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaHubert Robitaille - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaMartine Dunnwald - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaDong Hyun Kim - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaRoxane Pouliot - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaJulie Fradette - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, CanadaLucie Germain - LOEX Centre of Université Laval, Tissue Engineering and Regenerative Medicine: LOEX—FRQS Research Center of CHU de Québec and Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC, Canada
- Resource Type
- Journal article
- Publication Details
- Tissue engineering. Part A, Vol.19(7-8), pp.1023-1038
- DOI
- 10.1089/ten.tea.2012.0117
- PMID
- 23173810
- NLM abbreviation
- Tissue Eng Part A
- ISSN
- 1937-3341
- eISSN
- 1937-335X
- Language
- English
- Date published
- 04/2013
- Academic Unit
- Anatomy and Cell Biology; Craniofacial Anomalies Research Center
- Record Identifier
- 9984025304502771
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