title: Bioengineering a human plasma-based epidermal substitute with efficient grafting capacity and high content in clonogenic cells.
abstract: Cultured epithelial autografts (CEAs) produced from a small, healthy skin biopsy represent a lifesaving surgical technique in cases of full-thickness skin burn covering >50% of total body surface area. CEAs also present numerous drawbacks, among them the use of animal proteins and cells, the high fragility of keratinocyte sheets, and the immaturity of the dermal-epidermal junction, leading to heavy cosmetic and functional sequelae. To overcome these weaknesses, we developed a human plasma-based epidermal substitute (hPBES) for epidermal coverage in cases of massive burn, as an alternative to traditional CEA, and set up critical quality controls for preclinical and clinical studies. In this study, phenotypical analyses in conjunction with functional assays (clonal analysis, long-term culture, or in vivo graft) showed that our new substitute fulfills the biological requirements for epidermal regeneration. hPBES keratinocytes showed high potential for cell proliferation and subsequent differentiation similar to healthy skin compared with a well-known reference material, as ascertained by a combination of quality controls. This work highlights the importance of integrating relevant multiparameter quality controls into the bioengineering of new skin substitutes before they reach clinical development.
  
  
  CONCLUSIONS
  This work involves the development of a new bioengineered epidermal substitute with pertinent functional quality controls. The novelty of this work is based on this quality approach.
authors.affiliation: Biomedical Research Institute of French Armies, INSERM U1197, Clamart, France; Celogos, Paris, France; Alternative Energies and Atomic Energy Commission, Institute of Cellular and Molecular Radiobiology, Laboratory of Genomics and Radiobiology of Keratinopoiesis, INSERM UMR 967, Evry, France; Burn Treatment Unit, Percy Hospital, Clamart, France; Plastic Surgery Department, Percy Hospital, Clamart, France.
authors.forename: Maia M
authors.initials: MM
authors.lastname: Alexaline
authors.affiliation: Biomedical Research Institute of French Armies, INSERM U1197, Clamart, France; Celogos, Paris, France; Alternative Energies and Atomic Energy Commission, Institute of Cellular and Molecular Radiobiology, Laboratory of Genomics and Radiobiology of Keratinopoiesis, INSERM UMR 967, Evry, France; Burn Treatment Unit, Percy Hospital, Clamart, France; Plastic Surgery Department, Percy Hospital, Clamart, France.
authors.forename: Marina
authors.initials: M
authors.lastname: Trouillas
authors.affiliation: Biomedical Research Institute of French Armies, INSERM U1197, Clamart, France; Celogos, Paris, France; Alternative Energies and Atomic Energy Commission, Institute of Cellular and Molecular Radiobiology, Laboratory of Genomics and Radiobiology of Keratinopoiesis, INSERM UMR 967, Evry, France; Burn Treatment Unit, Percy Hospital, Clamart, France; Plastic Surgery Department, Percy Hospital, Clamart, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Nivet
authors.affiliation: Biomedical Research Institute of French Armies, INSERM U1197, Clamart, France; Celogos, Paris, France; Alternative Energies and Atomic Energy Commission, Institute of Cellular and Molecular Radiobiology, Laboratory of Genomics and Radiobiology of Keratinopoiesis, INSERM UMR 967, Evry, France; Burn Treatment Unit, Percy Hospital, Clamart, France; Plastic Surgery Department, Percy Hospital, Clamart, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Bourreau
authors.affiliation: Biomedical Research Institute of French Armies, INSERM U1197, Clamart, France; Celogos, Paris, France; Alternative Energies and Atomic Energy Commission, Institute of Cellular and Molecular Radiobiology, Laboratory of Genomics and Radiobiology of Keratinopoiesis, INSERM UMR 967, Evry, France; Burn Treatment Unit, Percy Hospital, Clamart, France; Plastic Surgery Department, Percy Hospital, Clamart, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Leclerc
authors.affiliation: Biomedical Research Institute of French Armies, INSERM U1197, Clamart, France; Celogos, Paris, France; Alternative Energies and Atomic Energy Commission, Institute of Cellular and Molecular Radiobiology, Laboratory of Genomics and Radiobiology of Keratinopoiesis, INSERM UMR 967, Evry, France; Burn Treatment Unit, Percy Hospital, Clamart, France; Plastic Surgery Department, Percy Hospital, Clamart, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Duhamel
authors.affiliation: Biomedical Research Institute of French Armies, INSERM U1197, Clamart, France; Celogos, Paris, France; Alternative Energies and Atomic Energy Commission, Institute of Cellular and Molecular Radiobiology, Laboratory of Genomics and Radiobiology of Keratinopoiesis, INSERM UMR 967, Evry, France; Burn Treatment Unit, Percy Hospital, Clamart, France; Plastic Surgery Department, Percy Hospital, Clamart, France.
authors.forename: Michele T
authors.initials: MT
authors.lastname: Martin
authors.affiliation: Biomedical Research Institute of French Armies, INSERM U1197, Clamart, France; Celogos, Paris, France; Alternative Energies and Atomic Energy Commission, Institute of Cellular and Molecular Radiobiology, Laboratory of Genomics and Radiobiology of Keratinopoiesis, INSERM UMR 967, Evry, France; Burn Treatment Unit, Percy Hospital, Clamart, France; Plastic Surgery Department, Percy Hospital, Clamart, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Doucet
authors.affiliation: Biomedical Research Institute of French Armies, INSERM U1197, Clamart, France; Celogos, Paris, France; Alternative Energies and Atomic Energy Commission, Institute of Cellular and Molecular Radiobiology, Laboratory of Genomics and Radiobiology of Keratinopoiesis, INSERM UMR 967, Evry, France; Burn Treatment Unit, Percy Hospital, Clamart, France; Plastic Surgery Department, Percy Hospital, Clamart, France.
authors.forename: Nicolas O
authors.initials: NO
authors.lastname: Fortunel
authors.affiliation: Biomedical Research Institute of French Armies, INSERM U1197, Clamart, France; Celogos, Paris, France; Alternative Energies and Atomic Energy Commission, Institute of Cellular and Molecular Radiobiology, Laboratory of Genomics and Radiobiology of Keratinopoiesis, INSERM UMR 967, Evry, France; Burn Treatment Unit, Percy Hospital, Clamart, France; Plastic Surgery Department, Percy Hospital, Clamart, France jjlataillade@gmail.com.
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Lataillade
mesh_terms: D000818:Animals; D002056:Burns / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D049109:Cell Proliferation; D020405:Dermis / Q000378:metabolism / Q000473:pathology; D004817:Epidermis / Q000378:metabolism / Q000473:pathology; D005260:Female; D064593:Heterografts; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism* / Q000473:pathology / Q000637:transplantation*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D010949:Plasma; D017768:Skin, Artificial
keywords: Artificial; Burns; Fibrin; Keratinocytes; Quality Control; Skin; Tissue Engineering
references.citation: Stem Cells Transl Med. 2013 Nov;2(11):830-8
references.pmid: 24101670
references.citation: J Clin Invest. 2014 Jul;124(7):3121-8
references.pmid: 24911148
references.citation: Burns. 2013 Jun;39(4):705-13
references.pmid: 23265935
references.citation: Cell Tissue Bank. 2006;7(1):47-53
references.pmid: 16511664
references.citation: Pediatr Surg Int. 1997 Sep;12(7):478-83
references.pmid: 9238111
references.citation: J Clin Invest. 2001 Jan;107(2):199-206
references.pmid: 11160136
references.citation: Burns. 2011 Jun;37(4):580-9
references.pmid: 21255936
references.citation: Nat Protoc. 2013 Feb;8(2):302-9
references.pmid: 23329003
references.citation: J Dermatol Sci. 2011 Jan;61(1):7-13
references.pmid: 21167690
references.citation: Nature. 2012 Sep 13;489(7415):257-62
references.pmid: 22940863
references.citation: Crit Care. 2010;14(5):R188
references.pmid: 20958968
references.citation: FASEB J. 2010 Jun;24(6):1838-51
references.pmid: 20097873
references.citation: Stem Cells. 2011 Aug;29(8):1256-68
references.pmid: 21674699
references.citation: Cytokine. 2013 Apr;62(1):1-21
references.pmid: 23490414
references.citation: J Invest Dermatol. 2004 May;122(5):1310-20
references.pmid: 15140236
references.citation: J Cell Biol. 1992 May;117(3):679-85
references.pmid: 1572896
references.citation: Burns. 2007 Jun;33(4):405-13
references.pmid: 17400392
references.citation: Surg Clin North Am. 1976 Apr;56(2):477-94
references.pmid: 772845
references.citation: J Burn Care Res. 2010 Jul-Aug;31(4):559-68
references.pmid: 20616650
references.citation: J Invest Dermatol. 2009 Feb;129(2):480-90
references.pmid: 18719609
references.citation: Burns. 1998 Nov;24(7):621-30
references.pmid: 9882060
references.citation: J Tissue Eng Regen Med. 2013 Jun;7(6):479-90
references.pmid: 22294482
references.citation: Ann Chir Plast Esthet. 2001 Dec;46(6):599-606
references.pmid: 11826710
references.citation: Trends Biotechnol. 1998 Jun;16(6):246-9
references.pmid: 9652135
references.citation: Burns. 2008 Jun;34 Suppl 1:S1-55
references.pmid: 18639739
references.citation: Exp Dermatol. 2010 Apr;19(4):387-92
references.pmid: 20201955
references.citation: Histochem Cell Biol. 2015 Jan;143(1):45-57
references.pmid: 25142512
references.citation: BMJ. 2004 Jul 17;329(7458):158-60
references.pmid: 15258073
references.citation: Stem Cells Transl Med. 2013 Jul;2(7):545-51
references.pmid: 23734060
references.citation: Burns. 2011 Nov;37(7):1087-100
references.pmid: 21802856
references.citation: Burns. 1991 Jun;17(3):181-4
references.pmid: 1892546
references.citation: Adv Skin Wound Care. 2007 Sep;20(9 Pt 1):493-508; quiz 509-10
references.pmid: 17762218
references.citation: Br J Dermatol. 2004 Nov;151(5):961-70
references.pmid: 15541073
references.citation: J Burn Care Rehabil. 1992 Jan-Feb;13(1):174-80
references.pmid: 1572850
references.citation: Int Wound J. 2013 Feb;10(1):6-12
references.pmid: 22958654
references.citation: Exp Cell Res. 2001 Nov 1;270(2):199-213
references.pmid: 11640884
references.citation: Br J Haematol. 2002 Jun;117(4):924-34
references.pmid: 12060132
references.citation: Transplantation. 1999 Sep 27;68(6):868-79
references.pmid: 10515389
references.citation: Cell. 1975 Nov;6(3):331-43
references.pmid: 1052771
references.citation: Transplantation. 2000 Dec 15;70(11):1588-98
references.pmid: 11152220
references.citation: Burns. 2006 Jun;32(4):395-401
references.pmid: 16621307
references.citation: J Am Coll Surg. 2013 Sep;217(3):533-55
references.pmid: 23816384
references.citation: J Cell Sci. 1996 May;109 ( Pt 5):1017-28
references.pmid: 8743949
references.citation: Proc Natl Acad Sci U S A. 1987 Apr;84(8):2302-6
references.pmid: 2436229
references.citation: Plast Reconstr Surg. 2001 Jan;107(1):105-15
references.pmid: 11176608
references.citation: Mol Med Today. 1999 May;5(5):201-8
references.pmid: 10322312
references.citation: Burns. 2014 Feb;40(1):82-8
references.pmid: 23764150
references.citation: Matrix Biol. 2008 Apr;27(3):163-70
references.pmid: 18029161
references.citation: J Skin Cancer. 2011;2011:936546
references.pmid: 21188234
references.citation: Hum Gene Ther. 1998 Mar 1;9(4):529-39
references.pmid: 9525314
references.citation: J Invest Dermatol. 2003 Jun;120(6):923-31
references.pmid: 12787116
references.citation: Burns. 2007 May;33(3):355-63
references.pmid: 17321690
references.citation: Eur J Dermatol. 2011 May;21 Suppl 2:4-11
references.pmid: 21628124
references.citation: Wound Repair Regen. 2008 Sep-Oct;16(5):585-601
references.pmid: 19128254
references.citation: J Cell Physiol. 2005 Nov;205(2):228-36
references.pmid: 15887229
references.citation: J Invest Dermatol. 1999 Jul;113(1):38-42
references.pmid: 10417616
references.citation: Burns. 2011 Sep;37(6):964-72
references.pmid: 21550174