Characterisation of the cell suspension harvested from the dermal epidermal junction using a ReCell® kit

被引:106
作者
Wood, Fiona M. [1 ,2 ,3 ]
Giles, Natalie [1 ,2 ]
Stevenson, Andrew [1 ]
Rea, Suzanne [1 ,3 ]
Fear, Mark [1 ,2 ,4 ]
机构
[1] Univ Western Australia, Burn Injury Res Unit, Sch Surg, Crawley, WA 6009, Australia
[2] Royal Perth Hosp, McComb Fdn, Perth, WA, Australia
[3] Princess Margaret Hosp, Perth, WA, Australia
[4] Univ Notre Dame Australia, Sch Med, Fremantle, WA, Australia
关键词
ReCell (R); Cells; Burn; Keratinocytes; Melanocytes; CULTURED EPITHELIAL AUTOGRAFT; HUMAN KERATINOCYTE ISOLATION; STEM-CELLS; IDENTIFICATION; POPULATION; VITILIGO; COVERAGE; GRAFTS; WOUNDS; MODEL;
D O I
10.1016/j.burns.2011.03.001
中图分类号
R4 [临床医学];
学科分类号
1002 ; 100602 ;
摘要
Background: The use of non-cultured autologous cells to promote wound healing and in reconstructive procedures is increasing. One common method for preparing these cells is the use of the ReCell (R) device. However, despite its current clinical use, no characterisation of the cell suspension produced using a ReCell (R)) device has been published. Objective: To characterise the Re Cell suspension that is applied to wounds for cell type, viability, yield, stability and proliferative potential. Methods: The ReCell (R) device was used to harvest cells from a 2 cm(2) piece of split-thickness skin isolated using a dermatome. The resulting cell suspension was analysed for cell yield, cell type, viability over time, proliferative potential and reproducibility. Results: Average viable cell yield was 1.7 x 10(6)/cm(2) of tissue, with 75.5% of the total cell isolate viable. Total viable cell number was not significantly reduced after 4 h storage at 22 degrees C or 4 degrees C, and was stable for 24 h at 4 degrees C. Proliferative potential was assessed using a colony forming assay, with 0.3% of viable cells isolated forming keratinocyte colonies. Predominantly the suspension contained keratinocytes (64.3 +/- 28.8%) and fibroblasts (30.3 +/- 14.0%), with a small population of melanocytes also identified (3.5 +/- 0.5%). Finally, the supernatant contained low total protein (0.92 mg/ml) and the supernatant had no significant effects on cell viability or growth when applied ex vivo. Conclusions: These results suggest the ReCell (R) device provides a method for the preparation of a cell suspension with high viability and proliferative potential, containing viable melanocytes and no apparent toxic cell debris. Further work on the sustained viability of these cells in vivo, and in particular after application to the wound, will be important to better understand the potential of the ReCell (R) device in the clinic. Crown Copyright (C) 2011 Published by Elsevier Ltd and ISBI. All rights reserved.
引用
收藏
页码:44 / 51
页数:8
相关论文
共 30 条
  • [1] FLOW CYTOMETRIC IDENTIFICATION OF PROLIFERATIVE SUBPOPULATIONS WITHIN NORMAL HUMAN EPIDERMIS AND THE LOCALIZATION OF THE PRIMARY HYPERPROLIFERATIVE POPULATION IN PSORIASIS
    BATACSORGO, Z
    HAMMERBERG, C
    VOORHEES, JJ
    COOPER, KD
    [J]. JOURNAL OF EXPERIMENTAL MEDICINE, 1993, 178 (04) : 1271 - 1281
  • [2] MONOCLONAL-ANTIBODIES FOR EPIDERMAL POPULATION ANALYSIS
    BAUER, FW
    BOEZEMAN, JBM
    VONENGELEN, L
    DEGROOD, RM
    RAMAEKERS, FCS
    [J]. JOURNAL OF INVESTIGATIVE DERMATOLOGY, 1986, 87 (01) : 72 - 75
  • [3] NORMAL KERATINIZATION IN A SPONTANEOUSLY IMMORTALIZED ANEUPLOID HUMAN KERATINOCYTE CELL-LINE
    BOUKAMP, P
    PETRUSSEVSKA, RT
    BREITKREUTZ, D
    HORNUNG, J
    MARKHAM, A
    FUSENIG, NE
    [J]. JOURNAL OF CELL BIOLOGY, 1988, 106 (03) : 761 - 771
  • [4] Comparison of cultured and uncultured keratinocytes seeded into a collagen-GAG matrix for skin replacements
    Butler, CE
    Yannas, IV
    Compton, CC
    Correia, CA
    Orgill, DP
    [J]. BRITISH JOURNAL OF PLASTIC SURGERY, 1999, 52 (02): : 127 - 132
  • [5] Cultured epithelial autografts in extensive burn coverage of severely traumatized patients: a five year single-center experience with 30 patients
    Carsin, H
    Ainaud, P
    Le Bever, H
    Rives, JM
    Lakhel, A
    Stephanazzi, J
    Lambert, F
    Perrot, J
    [J]. BURNS, 2000, 26 (04) : 379 - 387
  • [6] Use of a novel autologous cell-harvesting device to promote epithelialization and enhance appropriate pigmentation in scar reconstruction
    Cervelli, V.
    De Angelis, B.
    Spallone, D.
    Lucarini, L.
    Arpino, A.
    Balzani, A.
    [J]. CLINICAL AND EXPERIMENTAL DERMATOLOGY, 2010, 35 (07) : 776 - 780
  • [7] Cervelli V, 2009, ACTA DERMATOVENER CR, V17, P273
  • [8] CLUGSTON P A, 1991, Journal of Burn Care and Rehabilitation, V12, P533, DOI 10.1097/00004630-199111000-00008
  • [9] Melanocytes in cultured epithelial grafts are depleted with serial subcultivation and cryopreservation: Implications for clinical outcome
    Compton, CC
    Warland, G
    Kratz, G
    [J]. JOURNAL OF BURN CARE & REHABILITATION, 1998, 19 (04): : 330 - 336
  • [10] A porcine deep dermal partial thickness burn model with hypertrophic scarring
    Cuttle, Leila
    Kempf, Margit
    Phillips, Gael E.
    Mill, Julie
    Hayes, Mark T.
    Fraser, John F.
    Wang, Xue-Qing
    Kimble, Roy M.
    [J]. BURNS, 2006, 32 (07) : 806 - 820