+

MODEL

Journal of Plastic, Reconstructive & Aesthetic Surgery (2013) xx, 1e8

Practice of split-thickness skin graft storage and

histological assessment of tissue quality
Alicia Knapik a,d, Kai Kornmann a,d, Katrin Kerl b, Maurizio Calcagni a,
Claudio Contaldo a, Brigitte Vollmar c, Pietro Giovanoli a,
Nicole Lindenblatt a,*
a

Division of Plastic and Reconstructive Surgery, Department of Surgery, University Hospital Zurich, Raemistrasse 100, 8091
Zurich, Switzerland
b
Division of Dermatology, University Hospital Zurich, Raemistrasse 100, 8091 Zurich, Switzerland
c
Institute for Experimental Surgery, University of Rostock, Schillingallee 70, 18055 Rostock, Germany
Received 22 August 2012; accepted 11 February 2013

KEYWORDS
Skin graft;
Storage;
Cell viability;
Survey;
Burn

Summary Storage of split-thickness skin grafts (STSGs) represents a standard procedure in

burn surgery. The purpose of this study was to evaluate clinical routine of STSG preservation.
Further, we aimed at investigating the effect of storage on tissue integrity and cell viability,
proliferation, apoptosis and vascularization. A survey was performed among plastic surgery
centres in Europe. STSGs were harvested from healthy patients and analysed by histology
(HE, Verhoeffs, Massons Trichrome, Sirius Red) and immunohistochemistry (Ki67, TUNEL,
CD31). Cell viability was determined by MTT assay. The survey revealed that storage of STSGs
up to 10 days is common practice. STSGs mostly were stored at 4
C in saline-moisturized
gauze. Histology showed no disintegration of the tissue or a decrease of collagen and elastic
fibres. Proliferation increased to 22.5% of total cells after 3 days. On day 7 of STSG storage
apoptotic cells amounted for 25% of total cells. Cell viability decreased by 50% after day 3
of storage. Even though reportedly superior methods for skin grafts storage exist, most study
participants applied the simplest method of storage. Our data underscore this practice. However, a reduced cell viability after 3 days of storage may have an influence on graft healing.
2013 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by
Elsevier Ltd. All rights reserved.

* Corresponding author. Tel.: 41 44 255 1111; fax: 41 44 255 8977.

E-mail address: niclindenblatt@hotmail.com (N. Lindenblatt).
d
Equal contribution as first authors.
1748-6815/$ - see front matter 2013 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.bjps.2013.02.003

Please cite this article in press as: Knapik A, et al., Practice of split-thickness skin graft storage and histological assessment of tissue
quality, Journal of Plastic, Reconstructive & Aesthetic Surgery (2013), http://dx.doi.org/10.1016/j.bjps.2013.02.003

MODEL

A. Knapik et al.

Introduction
Storage of split-thickness skin grafts (STSGs) represents a
common practice in burn surgery to make use of surplus
harvested skin. Preservation in the refrigerator at 4e6
C in
moist saline gauze is the standard method. Other media,
e.g. Roswell Park Memorial Institute solution, University of
Wisconsin solution and Histidine-tryptophan-ketoglutarate
solution are rarely used,1,2 even though several authors have
reported their advantages over saline solution.3e6 Several
studies have focused on skin viability after storage in
different media,1,7 at different temperatures8 and for
different time periods.9 A reduction of tissue viability
generally was noted over time under all storage conditions.
The viability changes were found to be related to multiple
factors including storage solution, temperature, and graft
format.6 Most of these studies analysed the viability of the
preserved skin by histology and concentrated on the evaluation of skin integrity and cellular changes in the epidermis
and the epidermaledermal junction. Their focus was either
to enhance the storage conditions or just to understand in
general the changes which occur during storage. Despite
these facts, no study up to date has focused on the routine
of STSG short-term storage in a clinical setting.
Therefore, the present work has been designed to gain
insights into clinical routine concerning the storage and
application of human STSGs. Further, it was the goal of this
paper to evaluate the effect of storage on STSG integrity and
cell viability, cell proliferation, apoptosis and vascularity.

Materials and methods

On-line survey
An anonymous online survey was conducted among residents, senior surgeons and chief plastic surgeons in
Switzerland, Germany, Austria, Great Britain and France.
The questionnaire was created using survey monkey
(http://www.surveymonkey.com). It included 19 questions
concerning STSG storage conditions and clinical use. The
detailed questionnaire can be viewed as an online content.

Patients and STSG harvest

The study was approved by the local ethics committee.
Written and informed consent was given by every skin
Table 1

donor (n Z 17). Healthy patients were between 28 and 71

years old. Patients underwent different procedures of
dermolipectomy (abdominoplasty, belt lipectomy and
thigh lift). STSGs of 300 mm thickness were taken with an
air driven dermatome (Zimmer, Mu
nsingen, Switzerland)
and directly evaluated or stored for 3, 5 or 7 days in salinemoisturized gauze at 4
C in a refrigerator. In total skin
from 17 patients was analysed. The following groups were
established depending on the storage time: fresh skin
(n Z 5); 3 day storage (n Z 5), 5 day storage (n Z 5) 7 day
storage (n Z 5) and 14 day storage (n Z 4). Some skin
samples from the same patient were used after different
storage periods.

Histopathological evaluation of tissue integrity

Skin tissues were fixed in 4% formalin and subsequently
embedded in paraffin, according to standard procedures.
Sections (4 mm) were stained with Sirius Red for collagen
quantification, with a combination of Verhoeffs elastic and
Massons Trichrome for collagen and elastin fibers qualitative examination and with haematoxylin-eosin (HE) staining
for general morphological aspect.10 Samples were scored
concerning microscopic and macroscopic appearance by a
dermatopathologist in a blinded fashion. The score was
adapted from Ben-Bassat et al., Basaran et al. and TurhanHaktanir et al. (Table 1)1,7,11 Collagen quantification was
based on five random pictures taken with an AxioScope HRC
(Zeiss, Feldbach, Switzerland) and a 40 dry objective.
Pictures were further quantified with the analysis D software (Olympus Biosystems, Mu
nster, Germany) based on a
phase-contrast analysis and normalized to the total
surface.

Histological assessment of capillary density

Paraffin sections (4 mm) were stained with mouse antihuman CD31 (Dako, Glostrup, Denmark) and visualized
using diamino-benzidine (Dako, Glostrup, Denmark) as a
chromogen. Slides were scanned with the Mirax Midi Slide
Scanner (Zeiss, Feldbach, Switzerland) under a 20 dry
objective. Pictures were taken with a 3CCD colour camera
(1360  1024 pixels). Capillaries were counted in 5 random
dermal areas of a cross section of the skin and normalized
to the total dermal surface.

Parameters for microscopic and macroscopic assessment of graft quality after storage.

Microscopic parameter
Epidermal integrity
Epidermaledermal junction
Collagen organization
Apoptotic keratinocytes
& vacualisation
Macroscopic parameter
Colour
Pliability

Score
0
Destroyed
Destroyed
Amorph
Complete

1
Partial
Partial
Disturbed
Present

2
Normal
Normal
Normal
Normal

0
Blackened
Very rough

1
Mat grey
Minimally hardened

2
Normal
Pliable & smooth

Please cite this article in press as: Knapik A, et al., Practice of split-thickness skin graft storage and histological assessment of tissue
quality, Journal of Plastic, Reconstructive & Aesthetic Surgery (2013), http://dx.doi.org/10.1016/j.bjps.2013.02.003

MODEL

Practice of split-thickness skin graft storage

Immunohistochemical assessment of cell

proliferation
Paraffin embedded sections (4 mm) were single-labelled
with rabbit anti-human Ki67 (Abcam, Cambridge, UK) after
antigen-retrieval in boiling citrate buffer, followed by
antibody incubation and detection using ABC-Vectastain
(Reactolab, Servion, Switzerland). Quantification of stained
cells was performed by counting 10 representative fields for
each skin section using a 20 objective. The proliferation
rate is expressed as the ratio of the number of proliferating
cells divided by the total cell number.

Results
Survey participation
In total 158 questionnaires were sent out. 27 data sets were
returned and included for evaluation (response rate of
17.1%). The majority of participants were specialists
(22.2%), senior surgeons (55.6%) and chief surgeons (18.5%).
Almost all of the respondents worked in a University Hospital (96.3%). However, the major proportion of 70.4% was
employed in a division without a specialized burns unit.

Application of skin grafts

Evaluation of cell viability by MTT assay
A 3-(4,5)-dimethylthiazol-2,5-diphenyl tetrasolium bromide
(MTT) salt assay (Sigma, Buchs, Switzerland) was performed
in skin biopsies taken from fresh and stored skin as suggested by Ge et al.8 (n Z 6). Cell viability was expressed as
the ratio of its OD570 to its weight (mg) and normalized to
the 100% viability of fresh skin. The test was performed in
triplicates.

Determination of cell apoptosis rate

Apoptosis was assessed by in-situ Tdt-mediated dUTPbiotin nick-end labelling (TUNEL) using a commercially
available kit (Qbiogene, Illkirch, France). Quantification of
stained cells was performed by counting 10 representative
fields for each skin section using a 20 objective; the
apoptotic cell rate is given as proportion of the number of
positive cells/total number of cells.

Statistical analysis
Differences between values were assessed using 1-wayANOVA or KruskalleWallis tests followed by the appropriate
post-hoc comparison tests (Holm-Sidak and Dunns test,
respectively). All data were expressed as means  SE and
overall statistical significance was set at p < 0.05.

STSG represented a common method for wound coverage.

Approximately 70% of the respondents performed 50-200
STSG procedures per year. The most commonly used STSGs
were thin grafts of 200e300 mm (54.2%) and medium-thick
grafts of 300e400 mm (37.5%). Most wounds were covered
primarily with thin grafts (Figure 1). Ultrathin and thick
grafts were rarely used. All respondents applied STSGs on
acute wounds (tumour resections, etc.) as well as on
chronic wounds (leg ulcers, etc.), free/pedicled muscle
flaps, secondary wound closure (donor site after flap
elevation, etc.) and burn wound coverage.

Skin graft storage

Storage of STSGs after harvest was common, in 64% of the
hospitals STSGs were preserved for further use after harvest. In most units STSGs were wrapped in salinemoisturized gauze and kept at 4
C in a refrigerator,
rarely other media were used (Figure 2A and B). Despite
this practice 50% of the respondents had the opinion that
the quality of the graft deteriorates during storage. Fewer
than 20% of the respondents observed no decrease in
quality, and over 25% of the specialists had no opinion
concerning this issue. 30% of participants based their
opinion on a blackened colour of the graft after storage,
26% found the graft to be hardened, the remainder did not
see any morphological change of the graft. The observations of the clinicians differed concerning the graft taking

Figure 1 Results generated from the online survey depicting indications for use of STSGs of different thickness. Most surgeons
preferred to apply STSGs of thin or medium thickness on both acute and chronic wounds.

Please cite this article in press as: Knapik A, et al., Practice of split-thickness skin graft storage and histological assessment of tissue
quality, Journal of Plastic, Reconstructive & Aesthetic Surgery (2013), http://dx.doi.org/10.1016/j.bjps.2013.02.003

MODEL

A. Knapik et al.

Figure 2 Results generated from the online survey concerning the storage of split-thickness skin grafts in the clinical setting. (A)
Storage conditions: Most participants stored STSGs at 4e6
C in the refrigerator is a moist piece of gauze. (B) Preservation media:
The most common medium was saline. (C) Average storage time: The majority of clinicians stored STSGs for more than 7 days. (D)
Preferred period of storage before transplantation: Despite longer storage time most surgeons prefer to use fresh STSGs.

rate of stored STSGs, 50% estimated the graft take rate

generally as diminished after storage. The reported storage
time in over 50% of the hospitals was more than 7 days
(Figure 2C), despite the fact that 95% of the surgeons
consider the transplantation of fresh skin as the best
method (Figure 2D). Nonetheless, only 20% of the respondents refused to store skin and 40% would store it for a
maximum of 10 days.

Skin graft proliferation, apoptosis and cell viability

Results for Ki67- and TUNEL-labelling are shown in
Figure 3A. The proliferation rate about doubled (22.5%)

after 3 days of storage in comparison to fresh skin (10.6%)

and decreased significantly after 7 days of storage
(p < 0.05). On the other hand, apoptosis decreased by 10%
within the first three days but afterwards increased to a
level of 25% (p > 0.05). Cell viability of the graft dropped
within the first three days of storage significantly to
approximately 52% reaching finally a value of 44.1% in
comparison with fresh skin until 14 days of storage
(Figure 3B).
Macro - and microscopic graft evaluation according to
Ben-Bassat et al. revealed no atrophy of the tissues.11 No
destruction of collagen and elastic fibres was detected
after skin graft storage over a time period of 7 days

Figure 3 Effect of human STSG storage at 4
C for up to 7 days on cellular apoptosis and proliferation. (A) Proliferation rate
reached 22.5% on day 3 of storage and decreased significantly after 7 days of storage. On the other hand, apoptosis decreased by
10% within the first three days but afterwards increased to a level of 25%. (B) Viability change of human STSG stored at 4
C. Cell
viability decreased significantly after 3 days of storage to 44.1% and remained at this level during the subsequent days. Values are
means  SE. *p < 0.05.
Please cite this article in press as: Knapik A, et al., Practice of split-thickness skin graft storage and histological assessment of tissue
quality, Journal of Plastic, Reconstructive & Aesthetic Surgery (2013), http://dx.doi.org/10.1016/j.bjps.2013.02.003

MODEL

Practice of split-thickness skin graft storage

(Figure 4A and B). Collagen mass ratio in the skin did not
change over the time of storage (Figure 4C). Tissue evaluation revealed no changes in total graft score over the time
period of 7 days (Figure 4D).

Effect of storage on graft vasculature

HE and CD31 staining showed no abnormalities of vascular
morphology and vessel density. Capillaries did not show
signs of degeneration. Neither thrombosis nor vascular
obliterations were observed (Figure 5). Moreover, capillary
quantification revealed no significant changes of capillary
density in fresh skin (15.6  7.6 capillaries/mm2) and skin,
that had been stored for 7 days (15.0  7.8 capillaries/
mm2).

Discussion
Skin graft storage and delayed application is a frequent
practice in plastic and burn surgery. The method for storage
that appears to be most cost effective and simple is
refrigeration at 4e6
C in moist saline gauze. Skin grafting
to cover superficial acute and chronic wounds traditionally
provides good results for patients with a variety of defects
and low donor site morbidity.12 However, little clinical data
concerning actual practice in preservation of skin grafts
exist. Despite the fact that many different electrolyte and
nutrient rich solutions (e.g. Roswell Park Memorial Institute
solution, University of Wisconsin solution, Hartmanns solution, Dulbeccos Modified Eagle Medium, keratinocyte
nutrient MCDB 153, Ready Mix tissue culture medium,

Figure 4 Macro- and microscopical characterization of preserved human STSGs. HE (A) and Verhoeffs elastic/Massons Trichrome
(B) staining for collagen and elastic fibers (magnification 200) after 0, 3, 5 and 7 days of storage. Samples showed no significant
difference. (C) Relative collagen content in STSGs (Sirius Red stain), which remained stable over a period of 7 days. (D) Total graft
score of STSGs stored at 4
C in a saline-soaked gauze showing no deterioration of tissue quality. Values are means  SE. *p < 0.05.

Please cite this article in press as: Knapik A, et al., Practice of split-thickness skin graft storage and histological assessment of tissue
quality, Journal of Plastic, Reconstructive & Aesthetic Surgery (2013), http://dx.doi.org/10.1016/j.bjps.2013.02.003

MODEL

A. Knapik et al.

Figure 5 Representative endothelial-cell staining (CD31) on paraffin sections of fresh and 7 days old skin (magnification 200).
Vessel appearance on day 0 and 7 was identical without thrombosis or changes of cell morphology.

Marshalls solution, McCoys 5A medium etc.) exist,6,8,13 it

can be concluded from the presented study that almost all
of the respondents used the simple technique of storage in
saline gauze at 4e6
C for preservation. Previous studies
reported that skin graft storage at 4
C in saline for over 4
weeks preserved over 50% of keratinocyte viability.1,14 In
other studies, skin grafts stored under similar conditions
lost about 90% of their viability after 10 days and no graft
survival after 5 days of storage was observed.4,15
The application of fresh skin for transplantation remains
the gold standard.16 Our data indicate that in Europe
preservation of excess skin grafts for a possible use at a
later date is routinely applied. Next to the widely used
method of storage in saline solution at 4
C other possibilities of storage were previously investigated e.g.: at the
donor site as described by Shepard or Ashbell, cryopreservation or the utilization of different storage media and
temperatures.8,17e19 Li et al. recommends a storage of
meshed split-thickness skin grafts at 4
C in a culture media
(Dulbeccos Modified Eagle Medium) instead of a saline solution and grafting within seven days because of a significant loss of keratinocyte growth potency.6 Ge et al. also
investigated different storage temperatures (4
C/25
C)
and storage media (saline solution/DMEM) with the result of
recommended storage at 4
C in DMEM for maximum 96 h.8
In our survey, the time of storage is more than 7 days in
over 50% of the institutions. 40% of participants felt that
storage up to 10 days is feasible. The only alternative solutions used in two cases were Ready Mix solution and
Hartmanns solution, all participants stored the graft at
4
C in a refrigerator. Sterne et al. investigated also the
difference of storing the graft as a meshed or sheet graft
and the difference of a rolled or flat graft at 4
C wrapped
in a saline dampened gauze. They recommend storage at
4
C as a rolled sheet to achieve the best viability. However,
this group observed severe changes of skin graft
morphology and integrity after 7 days of storage. In our
split-thickness skin samples none of these changes were
seen as judged by an experienced clinical dermal pathologist. Since it is not stated in the paper how thick the
investigated split-thickness skin grafts were, and yet
changes all over the dermis are described, an explanation
may be that these samples simply were thicker than our
grafts with 300 mm thickness.20
As shown in an Australian survey by Lyall and Sinclair,
and also in our case, the surgeons seem in general to be

satisfied with their preferred method without the need for

alternatives.21 In addition it may preferably be used due to
its practicality and inexpensiveness.3,20 However, 50% of
respondents felt that the graft take rate is somewhat less
ideal after storage. Interestingly, no specific conclusion was
drawn from this and despite this assumption most surgeons
continue to store skin grafts and reuse them. One explanation may be that only 8 surgeons were from a hospital
with a specialised burns unit and have extensive experience
with skin grafting of large wounds. However, many surgeons
in Europe are involved in the coverage of smaller burns up
to 15% TBS, which often are treated in the smaller peripheral hospitals and not referred to a specialised burns
unit.
Since the simple storage in saline gauze proved to be the
preferred method of storage among the respondents in our
survey, we further investigated the effect of storage on the
skin graft tissue in vitro. Histological evaluation by a dermatopathologist revealed no macroscopic and only minor
microscopic changes within the graft over the time period
of 7 days. The overall total graft score showed no deterioration of colour and pliability as well as epidermal integrity, no detachment of the epidermaledermal junction, no
abnormalities of collagen organization and no keratinocyte
vacuolization. Therefore it does not appear surprising that
in practice grafts are stored up to and even longer than 7
days. These results are conform with previously published
observations of Sterne et al., who did not observe macroscopic and only minor epidermal microscopic changes
within the first seven days of storage at 4
C.20 Therefore it
is striking that more than the half of the respondents
observed macroscopic differences and diminished quality of
stored skin. However, despite their view, most clinicians
continue to use saline-stored stored skin grafts. It has to be
speculated that this is the case due to still sufficient - even
if slightly diminished - clinical take rates of STSGs. In light
of this it also has to be questioned, whether the reported
superiority of many other more expensive storage media is
of any clinical interest.3e6
Rosenquist et al. compared graft viability of meshed
and unmeshed skin and reported that meshing does not
affect the viability of the graft, demonstrating that even
mechanical stress does not affect the cell viability in a
long-lasting manner.22 In our study the MTT assay showed a
decrease of cell viability to a level of w50% within the first
3 days of storage and stayed on this level with only minor

Please cite this article in press as: Knapik A, et al., Practice of split-thickness skin graft storage and histological assessment of tissue
quality, Journal of Plastic, Reconstructive & Aesthetic Surgery (2013), http://dx.doi.org/10.1016/j.bjps.2013.02.003

MODEL

Practice of split-thickness skin graft storage

changes until the final storage day (14 days). This result is
similar to previous studies of Castagnoli et al.23 and
slightly lower than the viability reported by Ge et al.8
Additional tests for collagen or elastic fiber changes as
well as traditional scoring of the graft quality did not
reveal noticeable changes within different time points of
storage, which is congruent with the results of previous
studies. However, proliferation was found to be slightly
increased within the stored graft within the first 3 days,
most likely due to the acute hypoxic condition. This is in
line with previous studies showing increased proliferation
and differentiation of cells under hypoxic conditions and
inhibition of tumor cell proliferation by HIF1-alpha
silencing.24,25 In parallel apoptosis within the graft
increased after day 5 to 25% of cells as a consequence of
tissue storage. Cell viability was found be significantly
decreased to 50% already after 3 days of storage. In line
with this a recent study6 showed a progressive decrease of
cell viability over time during storage at 4e8
C. Cell
culture media were better than saline and Hartmanns
solution in this study in maintaining the viability and
growth capability of skin cells.
Even though STSG storage is performed frequently, it is
of interest, that the respondents indicated fresh skin as the
best graft as well as they observed diminished quality and
graft take after storage. Therefore, the diminished viability
after storage may have an impact on the graft take and the
proliferative phase during wound healing.
In conclusion our study reveals that skin graft storage is a
widely used practice in burn surgery and that most surgeons
prefer the method of preservation in saline-moistened
gauze at 4e6
C. Even though they have the impression
that skin graft take rate may be diminished after storage
they continue to apply this method. Histological evaluation
showed no visible deterioration of tissue integrity and
vascularity. However, the skin graft exhibited a slight increase of apoptotic cells after 5 days and a decreased
proliferation rate. Cell viability was reduced to half after
day 3 until day 14. Whether these observations have
implication for the take rate, vascularization and wound
healing of STSGs will have to be studied further in an in vivo
setting. Even though skin represents a complex organ with
multiple functions it can easily be preserved for up to 10
days under certain conditions without major changes in its
functionality and appearance. This insight may be of relevance for researchers working on the development of theshelf skin substitutes. Further, our findings could be translated into new skin graft material designs, basing on
reversible hypometabolism leading to an extended cell
survival prior to revascularisation and establishment of
nutrient and oxygen supply.

Conflict of interest statement

The authors declare that they have no conflicts of interest.

Acknowledgements
This study was financially supported by the Swiss National
Science Foundation (SNF-Grant 310030_127366).

References
1. Basaran O, Ozdemir H, Kut A, et al. Effects of different preservation solutions on skin graft epidermal cell viability and
graft performance in a rat model. Burns 2006;32:423e9.
2. Titley OG, Cooper M, Thomas A, Hancock K. Stored skin stored
trouble? Br J Plast Surg 1994;47:24e9.
3. Matsuka K, Hata Y, Yano K, Ito O, Matsuda H. Epidermal cell
viability in rat skin preserved at 4 degrees C. Ann Plast Surg
1993;31:358e63.
4. Cram AE, Domayer M, Shelby J. Human skin storage techniques: a study utilizing a nude mouse recipient. J Trauma
1983;23:924e6.
5. Cram AE, Domayer MA, Scupham R. Preservation of human
skin: a study of two media using the athymic (nude) mouse
model. J Trauma 1985;25:128e30.
6. Li Z, Overend C, Maitz P, Kennedy P. Quality evaluation of
meshed split-thickness skin grafts stored at 4
C in isotonic
solutions and nutrient media by cell cultures. Burns 2012 Feb
29 [Epub ahead of print].
7. Turhan-Haktanr N, Dilek FH, Ko
ken G, Demir Y, Yilmaz G.
Evaluation of amniotic fluid as a skin graft storage media
compared with RPMI and saline. Burns 2011;37:652e5.
8. Ge L, Sun L, Chen J, et al. The viability change of pigskin
in vitro. Burns 2010;36:533e8.
9. Ben-Bassat H, Chaouat M, Zumai E, et al. The Israel National
Skin Bank: quality assurance and graft performance of stored
tissues. Cell Tissue Bank 2000;1:303e12.
10. OConnor WN, Valle S. A combination Verhoeffs elastic and
Massons trichrome stain for routine histology. Stain Technol
1982;57:207e10.
11. Ben-Bassat H, Chaouat M, Segal N, Zumai E, Wexler MR,
Eldad A. How long can cryopreserved skin be stored to maintain adequate graft performance? Burns 2001;27:425e31.
12. Mardini S, Agullo FJ, Salgado CJ, Rose V, Moran SL, Chen HC.
Delayed skin grafting utilizing autologous banked tissue. Ann
Plast Surg 2009;63:311e3.
13. Hiroko Y, Yukihiro U, Toshihiko Y, et al. Cryopreserved cultured
epidermal allografts achieved early closure of wounds and
reduced scar formation in deep partial-thickness burn wounds
(DDB) and split-thickness skin donor sites of pediatric patients.
Burns 2001;27:689e98.
14. Cetin C, Ko
se AA, Aral E, et al. The effects of saline and plasma
on skin graft keratinocyte viability. Br J Plast Surg 2000;53:
418e9.
15. Fahmy FS, Navsaria HA, Frame JD, Jones CR, Leigh IM. Skin
graft storage and keratinocyte viability. Br J Plast Surg 1993;
46:292e5.
16. Leung JJ, Mus HB. Skin grafts. UTMJ 2009;2:86.
17. Shepard GH. The storage of split-skin grafts on their donor
sites. Plast Reconstr Surg 1972;49:115e22.
18. Ashbell TS. The storage of split-skin grafts on their donor sites
(letter). Plast Reconstr Surg 1972;50:178e9.
19. Kearney JN. Guidelines on processing and clinical use of skin
allografts. Clinica Dermatol 2005;23:357e64.
20. Sterne GD, Titley OG, Christie JL. A qualitative histological
assessment of various storage conditions on short term preservation of human split skin grafts. Br J Plast Surg 2000;53:
331e6.
21. Lyall PW, Sinclair SW. Australasian survey of split skin graft
donor site dressings. Aust N Z J Surg 2000;70:114e6.
22. Rosenquist MD, Kealey GP, Lewis RW, Cram AE. A comparison of
storage viability of nonmeshed and meshed skin at 4 degrees C.
J Burn Care Rehabil 1988;9:634e6.
23. Castagnoli C, Alotto D, Cambieri I, et al. Evaluation of donor
skin viability: fresh and cryopreserved skin using tetrazolioum
salt assay. Burns 2003;29:759e67.

Please cite this article in press as: Knapik A, et al., Practice of split-thickness skin graft storage and histological assessment of tissue
quality, Journal of Plastic, Reconstructive & Aesthetic Surgery (2013), http://dx.doi.org/10.1016/j.bjps.2013.02.003

MODEL

8
24. Valorani MG, Montelatici E, Germani A, et al. Pre-culturing
human adipose tissue mesenchymal stem cells under hypoxia
increases their adipogenic and osteogenic differentiation potentials. Cell Prolif 2012;45:225e38.

A. Knapik et al.
25. Shen SH, Kwan AL, Chen YY, Wang ZX. Effect of silencing
HIF-1a on proliferation, invasion and migration of glioblastoma U87 cells. Neurol Sci 2012 Mar 28 [Epub ahead of
print].

Please cite this article in press as: Knapik A, et al., Practice of split-thickness skin graft storage and histological assessment of tissue
quality, Journal of Plastic, Reconstructive & Aesthetic Surgery (2013), http://dx.doi.org/10.1016/j.bjps.2013.02.003