Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model
Aim. To study migration of syngeneic bone marrow MSCs and skin fibroblasts (FB), labeled by fluorochromes, after intravenous (IV) and local transplantation in a rat burn wound model (BWM). Methods. Rats were divided into 3 groups: C – without burn + IV injection of labeled MSCs and FBs mixture; O1 –...
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Інститут молекулярної біології і генетики НАН України
2015
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| Cite this: | Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model / E.A. Shchegelskaya, T.G. Grigorieva, E.A. Omelchenko, A.S. Zabirnyk, E.V. Markelova, S.G. Panibrattseva, I.A. Borovoy, G.I. Gubina-Vaculik, G.A. Oleynik // Вiopolymers and Cell. — 2015. — Т. 31, № 5. — С. 387-394. — Бібліогр.: 13 назв. — англ. |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1859942364602171392 |
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| author | Shchegelskaya, E.A. Grigorieva, T.G. Omelchenko, E.A. Zabirnyk, A.S. Markelova, E.V. Panibrattseva, S.G. Borovoy, I.A. Gubina-Vaculik, G.I. Oleynik, G.A. |
| author_facet | Shchegelskaya, E.A. Grigorieva, T.G. Omelchenko, E.A. Zabirnyk, A.S. Markelova, E.V. Panibrattseva, S.G. Borovoy, I.A. Gubina-Vaculik, G.I. Oleynik, G.A. |
| citation_txt | Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model / E.A. Shchegelskaya, T.G. Grigorieva, E.A. Omelchenko, A.S. Zabirnyk, E.V. Markelova, S.G. Panibrattseva, I.A. Borovoy, G.I. Gubina-Vaculik, G.A. Oleynik // Вiopolymers and Cell. — 2015. — Т. 31, № 5. — С. 387-394. — Бібліогр.: 13 назв. — англ. |
| collection | DSpace DC |
| container_title | Вiopolymers and Cell |
| description | Aim. To study migration of syngeneic bone marrow MSCs and skin fibroblasts (FB), labeled by fluorochromes, after intravenous (IV) and local transplantation in a rat burn wound model (BWM). Methods. Rats were divided into 3 groups: C – without burn + IV injection of labeled MSCs and FBs mixture; O1 – BWM + IV injection of labeled MSCs and FBs mixture; O2 – BWM + fibrin matrix filled with labeled cell mixture. MSCs were labeled by green fluorochrome, and fibroblasts – by red one. The presence of labeled cells in cryocuts of the skin, liver, kidney and bone marrow was assessed on the 3 and 7 days after transplantation. Results. Skin FBs selective migration to the regenerating burn wound and MSCs accumulation in the kidneys were found in rats of group O1 on day 7 after the IV injection. The labeled cells proliferated in the transplanted fibrin matrix and participated in the wound regeneration. MSCs partly migrated to the bone marrow after the IV injections. Conclusions. IV transplanted syngeneic bone marrow MSCs and skin FBs (passage 0) migrate to the burn wound and participate in the healing. Migration of bone marrow MSCs in the kidneys can prevent kidney failure after burn.
Мета. Вивчення міграції сингенних МСК кісткового мозку і фібробластів (ФБ) шкіри, мічених флюорохромами, після внутрішньовенної (ВВ) та локальної трансплантації у щурів з моделлю післяопікової рани (МПР). Методи. Щури були розділені на 3 групи: К– без опіку + ВВ введення мічених МСК і ФБ; О1 – МПР + ВВ мічених МСК і ФБ; О2 – МПР + фібринова матриця з сумішшю мічених МСК і ФБ. МСК фарбували зеленим флюорохромом, а фібробласти – червоним. Розподіл мічених клітин на кріозрізах шкіри, печінки, нирок і кісткового мозку щурів вивчали на 3 і 7 добу після трансплантації. Результати. Виявлена вибіркова міграція ФБ шкіри в післяопікову рану і накопичення МСК КМ в нирках тварин групи О1 на 7 добу після ВВ введення мічених МСК і ФБ. У трансплантованій на рану фібриновій матриці мічені клітини проліферували та брали участь у процесі регенерації рани. При ВВ введенні МСК КМ спостерігалася їх міграція в кістковий мозок. Висновки. Системно введені сингенні МСК КМ і ФБ шкіри (0 пасаж) мігрують в зону післяопікової рани і беруть участь в її загоєнні. Міграція МСК КМ в нирки може попередити ниркову недостатність, що розвивається після опіку.
Цель. Изучить миграции сингенных МСК костного мозга и фибробластов (ФБ) кожи, меченых флюорохромами, после их внутривенной (ВВ) и локальной трансплантации крысам с моделью послеожоговой раны (МПР). Методы. Крыс разделили на 3 группы: К – без ожога + ВВ введение смеси меченых МСК и ФБ; О1 – МПР + ВВ введение смеси меченых МСК и ФБ; О2 – МПР + фибриновая матрица с мечеными МСК и ФБ. МСК метили зеленым флюорохромом, фибробласты – красным. Наличие меченых клеток в коже, печени, почках и костном мозге крыс оценивали на 3 и 7 сутки после трансплантации на криосрезах и мазках. Результаты. Обнаружена избирательная миграция ФБ кожи в зону раны и накопление МСК КМ в почках крыс группы О1 на 7 сутки после ВВ введения меченых МСК и ФБ. В трансплантированной на рану фибриновой матрице меченые клетки активно делились и участвовали в ее регенерации. При ВВ введении МСК КМ частично мигрировали в костный мозг. Выводы. Системно трансплантированные сингенные МСК КМ и ФБ кожи 0 пассажа мигрируют в зону послеожоговой раны и участвуют в ее заживлении. Миграция МСК КМ в почки может предупредить вызванную ожогом почечную недостаточность.
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387
E. A. Shchegelskaya, T. G. Grigorieva, E. A. Omelchenko
© 2015 E. A. Shchegelskaya et al.; Published by the Institute of Molecular Biology and Genetics, NAS of Ukraine on behalf of Biopolymers and Cell.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/),
which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited
Biomedicine ISSN 0233-7657
Biopolymers and Cell. 2015. Vol. 31. N 5. P. 387–394
doi: http://dx.doi.org/10.7124/bc.0008FB
UDC 576.08:576.524:57.089.6
Migration of labeled bone marrow MSCs and skin fibroblasts
after systemic and local transplantation in rat burn wound model
E. A. Shchegelskaya2, T. G. Grigorieva1, E. A. Omelchenko1,
A. S. Zabirnyk1, E. V. Markelova1, S. G. Panibrattseva1, I. A. Borovoy3,
G. I. Gubina-Vaculik2, G. A. Oleynik1
1 Kharkiv Medical Academy of Postgraduate Education
58, Korchagintsev Str, Kharkiv, Ukraine, 61176
2 Kharkiv National Medical University
4, Lenina ave., Kharkiv, Ukraine, 61022
3 Institute for Scintillation Materials, NAS of Ukraine
60, Lenina ave., Kharkiv, Ukraine, 61001
shcheglik2@gmail.com
Aim. To study migration of syngeneic bone marrow MSCs and skin fibroblasts (FB), labeled by fluoro-
chromes, after intravenous (IV) and local transplantation in a rat burn wound model (BWM). Methods. Rats
were divided into 3 groups: C – without burn + IV injection of labeled MSCs and FBs mixture; O1 – BWM +
IV injection of labeled MSCs and FBs mixture; O2 – BWM + fibrin matrix filled with labeled cell mixture.
MSCs were labeled by green fluorochrome, and fibroblasts - by red one. The presence of labeled cells in cry-
ocuts of the skin, liver, kidney and bone marrow was assessed on the 3 and 7 days after transplantation.
Results. Skin FBs selective migration to the regenerating burn wound and MSCs accumulation in the kidneys
were found in rats of group O1 on day 7 after the IV injection. The labeled cells proliferated in the trans-
planted fibrin matrix and participated in the wound regeneration. MSCs partly migrated to the bone marrow
after the IV injections. Conclusions. IV transplanted syngeneic bone marrow MSCs and skin FBs (passage 0)
migrate to the burn wound and participate in the healing. Migration of bone marrow MSCs in the kidneys can
prevent kidney failure after burn.
K e y w o r d s: burn wound, migration, fibroblasts, MSCs, fluorochrome, transplantation.
Introduction
The transplantation of skin fibroblasts (FB) and
bone marrow mesenchymal stem cells (BM MSCs)
is successfully used in the treatment of deep and ex-
tensive burns in many modern burn centers of the
world. The beneficial effect of fibroblasts on the
process of healing burn wounds was proven 20
years ago, when transplantations of both auto- and
allofibroblasts on different matrices were found to
be efficient [1]. BM MSCs became a promising can-
didate for application in the regenerating medicine
due to their pluripotency. During the treatment of
the wounds, MSCs are transplanted into the latter
via injections or using biodegradable matrices,
based on collagen, fibrin, gelatin, chitosan, and hy-
aluronic acid [2]. The positive effect of BM MSCs
in regenerating skin at different stages of wound
healing was proven both in the experiments on ani-
mals and in the series of clinical trials, and it was
established that the injury stimulates the acceptance
of MSCs by skin and induces their differentiation
388
E. A. Shchegelskaya, T. G. Grigorieva, E. A. Omelchenko et al.
into skin cells [3]. One of possible mechanisms of
healing skin wounds is MSCs differentiation into
fibroblasts in the injured zone and their synthesis of
extracellular matrix proteins.
The comparative evaluation of the efficiency of
transplanting embryonic fibroblasts and bone mar-
row MSCs in the treatment of deep burns in rats
demonstrated that the transplantation of these cells
on the surface of the wound decreases its cellular in-
filtration, accelerates the formation of new vessels
and granulation tissue. Moreover, the MSCs trans-
plantation leads to a higher regeneration rate com-
pared to the transplantation of embryonic fibroblasts
[4]. The most efficient method of treating burn
wounds is a complex approach, which simulates nor-
mal skin structure and promotes its faster and more
normal formation. For instance, it was previously
demonstrated by us that the use of biodegradable fi-
brin films, covered with BM MSCs or FBs culture
for the treatment of wound surface of the burn model
in combination with xenoskin and autokeratinocytes
leads to the formation of the dermal type regenerate
and the 2–5-fold acceleration of wound healing
compared to the control [5].
There is still an unresolved issue of the efficiency
of IV application of BM MSCs and especially skin
FBs in the burn wound treatment and burn disease in
general, which is usually characterized by multiple
organ failure. One of the studies, comparing local
and systemic injections of cultivated dermal pluripo-
tent cells to rats with wounds in combination with
radiation injury or without one, demonstrated that
with local injection the effect was observed sooner,
but only systemic transplantation of cells had impact
on healing the wounds of radiation-exposed rats. As
the radiation damaged bone marrow (MSCs reserve),
externally introduced stromal cells took an active
part in the wound regeneration [6]. It is known that
the MSCs cultures of different passages and origin
are heterogeneous populations of cells, different in
their ability to migrate to one or another zone of
damage or inflammation. To study the mechanisms
of cell migration, MSCs are transfected with the
genes of fluorescent proteins or the cells are stained
with vital fluorochromes. These data are used to
elaborate the methods of targeted migration of MSCs
into a specific niche in the organism [7].
The study of the migration ways of MSCs and
skin FB, stained with fluorescent dyes, at their sys-
temic and local transplantation to rats with wound
models, may clarify the role of these cells in wound
regeneration.
The aim of this work was to study the migration
of BM MSCs and skin FB, labeled with green and
red vital fluorochromes respectively, after their in-
travenous (IV) and local syngeneic transplantation
in the fibrin matrix to rats with burn wound model
(BWM).
Materials and Methods
The study was conducted on 18 female rats of
Wistar line, with the weight of 250–300 g, which
were used to simulate a deep burn wound of 20 %
area in conditions of thiopental-induced anesthesia.
After local removal of skin hair on the back of the
animals, the latter were inflicted a contact burn using
a round tip of the electric soldering iron with the di-
ameter of 4 cm. Primary necrectomy was performed
on all the experimental animals 24 h later.
The rats were divided into three groups of 6 ani-
mals: C – no burn + IV injection of labeled MSCs
and FB; О1 – BWM + IV injection of labeled MSCs
and FB; О2 – BWM + fibrin matrix with the mixture
of labeled MSCs and FBs + xenoskin. Acellular pig
skin (Kombustiolog Company, Ternopil) was used
as xenoskin.
The localization of labeled cells in skin, liver, kid-
neys and bone marrow of rats was studied on days 3
and 7 after the transplantation.
The experiments were conducted in the vivarium
of the Kharkiv Medical Academy of Postgraduate
Education in accordance with the international re-
quirements of the European Convention for the
Protection of Vertebrate Animals (Strasbourg, 1985)
and General principles of conducting experiments
on animals (Kyiv, 2004). All the manipulations with
animals, including decapitation, were performed us-
ing thiopental anesthesia.
389
Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model
The methods of preparing the cultures of MSCs
and skin FBs and their labeling with fluores-
cent dyes
The methods, described by us previously [5],
were used to obtain primary cultures (passage 0) of
bone marrow MSCs and skin fibroblasts of rats. The
reagents, sera and media of SIGMA-ALDRICH
company (USA), and the culture dishes of PAA com-
pany (Austria) were used in the work.
The fluorescent dyes DIO C18 (3,3’-dioctadecyloxa-
carbocyanine bromide, C53H85BrN2O2, FW 863,16)
and Rhod Chol (cholesterol ester of rhodamine С,
C55H75ClN2O3, FW 847,65), synthesized in the
Institute of Scintillation Materials, NASU, have low
solubility in water and, due to their hydrophobic prop-
erty, are easily bound to lipid areas of membrane struc-
tures of the cells. This property allows using them as
intravital fluorescent markers for different types of cells.
The fluorescence maximum for DIO C18 with the
excitation in the range of 455–490 nm is 515 nm
(green fluorescence). The dye Rhod_Chol is remark-
able for red fluorescence (maximum 580 nm) at the
excitation of 490 nm.
These dyes are closely bound to the membrane
structures of cell cytoplasm, do not penetrate the nu-
cleus, are resistant to discoloration and well-kept in the
cells, are not cytotoxic in optimal conditions of dying.
The stock solutions of fluorochromes DiO С18
(λem=515 nm, green) and Rhod Chol (λem=580 nm,
red), prepared with 100 % dimethylsulfoxide
(DMSО, 10–3 М), were used to prepare their work-
ing solutions with saline solution (SS) and the final
concentration of the dye in the solution was 2х10–
5М. MSCs were labeled with green fluorochrome,
and fibroblasts – with red one. For this reason the
cell suspensions, obtained after the cultivation, were
washed off fetal serum using centrifugation in
Hanks’ solution, then the pellet was resuspended in
the working solutions of dyes (MSCs – in the solu-
tion DIO C18, FBs – in Rhod Chol solution). The
cells were incubated in the dye solutions for 15 min
at room temperature. Then the cells were washed in
SS off the excess of fluorochrome by centrifugation
at 430 g for 7 min. The pellet was resuspended in SS
and labeled MSCs and FBs were mixed in equal vol-
umes (400 thousand MSCs/ml, 400 thousand FB/
ml). The rats from groups C and O1 were given IV
injections of 0.5 ml of the mixture of labeled MSCs
and FB. Some labeled cells from this mixture were
plated in Petri dishes and cultivated for 3 and 7 days
for the control of the dying quality and cell viability.
The preparation of a fibrin matrix with cells
Labeled MSCs and FBs were resuspended in
Hanks’ solution (200 thousand cells per ml) and
mixed with blood plasma, 0.3 % calcium chloride
was added to the mixture, the portions of 2 ml were
poured into each Petri dish (d=35mm) and incubated
at 37°С for 15 min until fibrin gel was formed. In
Experiment 2 the fibrin matrices with enclosed la-
beled fibroblasts (red) and MSCs (green) were trans-
ferred to the surface of burn wound and covered with
acellular pig xenoskin to prevent wound drying.
Histological methods of analysis
The samples of skin, liver and kidneys with the
size of 2 cc were taken from rats on days 3 and 7
after the cell transplantation and transported on ice
to the laboratory, where 20–25 µm thick cryocuts
were made using a freezing microtome MK-25
(Russia). 10–15 cryocuts were placed on each slide.
100–120 cuts were prepared from each organ. Bone
marrow smears were made from the femur bone of
all the animals. The presence and distribution of flu-
orescent cells on cryocuts of tissues and bone mar-
row smears were estimated using a luminescent mi-
croscope Axioscope 40 (Carl Zeiss, Germany) with
the magnification of 100. Instudio program was used
to register microphotographs.
Statistical analysis
The comparative estimation of the number of fluo-
rescent red and green cells in the liver, kidneys and
skin of rats involved the determination of the average
number of labeled cells per one cut after they were
counted in all the cryocuts of each organ. The data
were presented as mean ± standard deviation (SD).
The statistical analysis was performed using Statsoft
390
E. A. Shchegelskaya, T. G. Grigorieva, E. A. Omelchenko et al.
Statistica 8.0 (license No. STA862D175437Q);
Student’s t-criterion was used for comparison. The
value of P < 0.05 was considered statistically reliable.
Results and Discussion
Fluorescent dyes DIO С18 (green) and Rhod Chol
(red) were previously used to study the migration of
BM MSCs to treat Parkinson-like syndrome in rats [8].
This study also demonstrated the MSCs affinity to the
zone of chemical destruction of brain in case of the
intravenous injection. The same fluorochromes were
used here to study the migration of primary cultures of
skin FB, labeled red, and BM MSCs, labeled with
green fluorochrome, in the organism of rats with a
burn wound model after a single intravenous injection.
It was established that labeled FBs and MSCs in
the mixed culture preserve their normal fibroblast-
like phenotype and proliferate actively; the cell cyto-
plasm manifests green fluorescence of MSCs and
red one – of fibroblasts (Fig. 1). Fibroblasts divide
more actively, therefore, 7 days later they are much
more numerous in the culture compared to MSCs.
The analysis of cryopreparations demonstrated that
single MSCs and FBs (2 cells per 6–8 cuts) were
found in all the studied organs of rats in group C (IV
injection of the mixture of MSCs and FBs to rats with-
out any burn) in different periods after the transplanta-
tion. In the bone marrow only single MSCs (3–4 cells
per smear) were found, and none FBs.
On day 7 after the IV injection of the cell mixture,
group O1 demonstrated a considerable (10-fold) in-
crease in the number of green MSCs in the kidneys
compared to the control (3–5 cells per each cut on
average) (Fig. 3 B) and a simultaneous multiple in-
crease in the number of red FBs in the burn wound
area (8–12 cells per each cut) (Fig. 3 A), compared to
the control and 3 days after the cell injection. A con-
siderable increase in the number of green MSCs (2–3
per each cut), compared to the control and day 3 after
IV injection in O1, was also found on the cuts of the
wound (Fig. 4 A, B). Single green MSCs were found
in the liver and bone marrow of animals of this group
on days 3 and 7 of the studies. Fibroblasts on the cuts
of kidneys and liver were observed very rarely.
The zone of inflammation in the area of spontane-
ously healing burn wound is likely to produce specific
factors which lead to the selective migration of skin
FBs into this target at their systemic injection. BM
MSCs also migrate into the same injured zone, but in a
smaller amount. Contrary to fibroblasts, a part of
MSCs also migrate into the liver, kidneys and bone
marrow, which may be explained by the fact that they
are less differentiated cells, expressing a wide spec-
trum of receptors, and may participate in the regenera-
tion of these organs and tissues, which undergo a toxic
burden due to the development of a burn disease.
As for the animals from group O2, red (FB) and
green (MSCs) fluorescent cells were found on days 3
(Fig. 5A) and 7 of the studies in the area of wound
surface, which was covered with a fibrin matrix with
enclosed labeled MSCs and FB. Moreover, on day 7
there were more labeled cells in the regeneration
zone than on day 3, which may testify to FBs and
MSCs entering the phase of active division. Similar
to the cells culture, FBs in the regenerating wound
divided more actively, therefore, they were 3–4 times
more abundant on the cuts compared to MSCs
(Fig. 5B). On day 7 it was evident by the morpho-
logical parameters that the wound healing in group
O2 was faster and of a higher quality than in O1.
Neither red (FB) nor green (MSCs) fluorescent cells
were found in the liver, kidneys and bone marrow of
these animals during the period of analysis. Therefore,
these cells cannot migrate into the mentioned organs
with this type of transplantation during this period.
Being enclosed into the fibrin matrix, they have di-
rect contact with the wound and take an active part in
the regenerating processes of its healing.
Single green MSCs in the amount of 5–10 cells
per smear were found in the preparations while
studying the bone marrow smears of the animals
from groups C and O1 (Fig. 6 A, B). Fibroblasts were
not found in the bone marrow of rats after the IV
injection. There were no reliable differences in the
amount of MSCs in bone marrow of control and ex-
periment animals. Thus, the phenomenon of cell
homing was confirmed for IV transplantation of BM
MSCs. Similar migration of MSCs into the bone
391
Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model
marrow, lungs and tumor was demonstrated for sys-
temic and local introduction of MSCs of adipose tis-
sue, labeled with fluorescent red protein TurboFP635
[9]. MSCs express many receptors and molecules of
cell adhesion, which promote the migration into the
bone marrow and the target tissue. Nonetheless, pre-
cise mechanisms of MSCs homing have not been
elucidated completely [10]. Single red fluorescent
cells were found on the cryocuts of intact animals
skin after IV injection of fibroblasts. In this case we
may assume the homing of skin fibroblasts, which
has been studied even less than MSCs homing.
Notably, the data, obtained by us, refer to the speci-Fig. 1. A mixed culture of labeled FBs (red) and MSCs (green)
on day 7 of in vitro cultivation. Luminescent microscopy. x200.
A B
Fig. 2. BM MSCs, labeled with DIO C18 (green), on the cryocut of kidney of the rat from group O1 on day 7 after IV injection of
the labeled cell mixture. A – luminescent, B – light microscopy. x100.
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Fig. 3. A number of labeled FBs (A) and MSCs (B) in the tissues of liver, kidneys, and skin of rats on day 7 after IV injection of the
labeled FBs and MSCs mixture. 1 – Control, 2 – Experiment 1; – liver, – kidney, – skin
392
E. A. Shchegelskaya, T. G. Grigorieva, E. A. Omelchenko et al.
A B
Fig. 6. BM MSCs, labeled with green fluorochrome in the smear of rat bone marrow from O1 on day 7 after IV injection of the la-
beled cell mixture. A – luminescent, B – light microscopy. x100.
A B
Fig. 5. Labeled FBs (red) and BM MSCs (green) on days 3 (A) and 7 (B) of the fibrin matrix transplantation into the wound on the
cut of skin of the regenerating wound of group O2 rats. Luminescent microscopy. x100.
A B
Fig. 4. Labeled FBs (red) and MSCs (green) on the cryocuts of the regenerating skin from the burn wound on day 7 after IV injection
of labeled cells. A – luminescent, B – light microscopy. x100.
393
Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model
ficities of cell migration of FB and BM MSC pri-
mary cultures at syngeneic transplantation. It is
known that after multiple subcultivations or cryo-
preservation MSCs and FBs may change the spec-
trum of expressed receptors and cytokines, and thus
their migration preferences may be different. The
application of allogeneic or xenogenic cell cultures
while studying the migration of these cells may also
have some specificities [11, 12].
The method of intravital staining of cells with
fluorochromes allows studying the ways of migra-
tion of labeled cells (especially early passages) in the
organism of animals in short terms (up to 2 weeks)
after the injection. The disadvantage of this method
is the fact that after the cell division the labeled
membrane structures are distributed among daughter
cells and the fluorescence gradually weakens. Cell
lines or MSC cultures, transfected with the genes of
fluorescent proteins, such as GFP, are more suitable
for long-term studies of cell migration [13].
The data, obtained by us, may be used for the de-
velopment of an optimal method of the cell therapy
while treating extensive and deep burn wounds. For
instance, both local transplantation of fibroblasts and
bone marrow MSCs and their IV injection may be
used for wound treatment. Systemic transplantation
of MSCs may be especially efficient in treating the
burn disease, because on day 7 after the burn they
migrate into kidneys which carry a considerable bur-
den in case of the extensive and deep burns.
Conclusions
1. The selective migration of skin FBs into the
area of spontaneously regenerating burn wound
and the accumulation of BM MSCs in the kidneys
of these animals were found on day 7 after IV injec-
tion of the mixture of syngeneic labeled MSCs
and FB.
2. The labeled cells (MSCs and FB) proliferate
and participate in the process of wound healing in
the fibrin matrix, transplanted onto the wound, more-
over, FBs divide faster than MSCs.
3. Regardless of the wound, the IV injected syn-
geneic BM MSCs migrate into their niche – bone
marrow. In similar conditions FBs do not migrate to
the bone marrow.
4. Simultaneous systemic transplantation of pri-
mary cultures of BM MSCs and skin FBs may be
efficient for healing the burn wounds and prevention
of the burn disease.
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Міграція мічених МСК кісткового мозку і фібробластів
шкіри після їх системної та місцевої трансплантації
у щурів з моделлю післяопікової рани
О. А. Щегельська, Т. Г. Григор’єва, О. А. Омельченко,
А. С. Забірник, О. В. Маркелова, С. Г. Панібратцева,
І. А. Боровий, Г. І. Губіна-Вакулик, Г. А. Олійник
Мета. Вивчення міграції сингенних МСК кісткового мозку і
фібробластів (ФБ) шкіри, мічених флюорохромами, після вну-
трішньовенної (ВВ) та локальної трансплантації у щурів з мо-
деллю післяопікової рани (МПР). Методи. Щури були розді-
лені на 3 групи: К – без опіку + ВВ введення мічених МСК і
ФБ; О1 – МПР + ВВ мічених МСК і ФБ; О2 – МПР + фібрино-
ва матриця з сумішшю мічених МСК і ФБ. МСК фарбували
зеленим флюорохромом, а фібробласти – червоним. Розподіл
мічених клітин на кріозрізах шкіри, печінки, нирок і кісткового
мозку щурів вивчали на 3 і 7 добу після трансплантації.
Результати. Виявлена вибіркова міграція ФБ шкіри в післяопі-
кову рану і накопичення МСК КМ в нирках тварин групи О1 на
7 добу після ВВ введення мічених МСК і ФБ. У транспланто-
ваній на рану фібриновій матриці мічені клітини проліферува-
ли та брали участь у процесі регенерації рани. При ВВ введен-
ні МСК КМ спостерігалася їх міграція в кістковий мозок.
Висновки. Системно введені сингенні МСК КМ і ФБ шкіри (0
пасаж) мігрують в зону післяопікової рани і беруть участь в її
загоєнні. Міграція МСК КМ в нирки може попередити ниркову
недостатність, що розвивається після опіку.
К л юч ов і с л ов а: післяопікова рана, міграція, фібробласти,
МСК, флюорохроми, трансплантація.
Миграция меченых МСК костного мозга
и фибробластов кожи после их системной и местной
трансплантации у крыс с моделью послеожоговой раны
Е. А. Щегельская, Т. Г. Григорьева, Е. А. Омельченко,
А. С. Забирник, Е. В. Маркелова, С. Г. Панибратцева,
И. А. Боровой, Г. И. Губина-Вакулик, Г. А. Олейник
Цель. Изучение миграции сингенных МСК костного мозга и
фибробластов (ФБ) кожи, меченых флюорохромами, после их
внутривенной (ВВ) и локальной трансплантации крысам с мо-
делью послеожоговой раны (МПР). Методы. Крыс разделили
на 3 группы: К – без ожога + ВВ введение смеси меченых МСК
и ФБ; О1 – МПР + ВВ введение смеси меченых МСК и ФБ;
О2 – МПР + фибриновая матрица с мечеными МСК и ФБ.
МСК метили зеленым флюорохромом, фибробласты – крас-
ным. Наличие меченых клеток в коже, печени, почках и кост-
ном мозге крыс оценивали на 3 и 7 сутки после транспланта-
ции на криосрезах и мазках. Результаты. Обнаружена избира-
тельная миграция ФБ кожи в зону раны и накопление МСК КМ
в почках крыс группы О1 на 7 сутки после ВВ введения мече-
ных МСК и ФБ. В трансплантированной на рану фибриновой
матрице меченые клетки активно делились и участвовали в ее
регенерации. При ВВ введении МСК КМ частично мигрирова-
ли в костный мозг. Выводы. Системно трансплантированные
сингенные МСК КМ и ФБ кожи 0 пассажа мигрируют в зону
послеожоговой раны и участвуют в ее заживлении. Миграция
МСК КМ в почки может предупредить вызванную ожогом по-
чечную недостаточность.
К л юч е в ы е с л ов а: ожоговая рана, миграция, фибробла-
сты, МСК, флюорохромы, трансплантация.
Received 28.07.2015
|
| id | nasplib_isofts_kiev_ua-123456789-152613 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 0233-7657 |
| language | English |
| last_indexed | 2025-12-07T16:12:22Z |
| publishDate | 2015 |
| publisher | Інститут молекулярної біології і генетики НАН України |
| record_format | dspace |
| spelling | Shchegelskaya, E.A. Grigorieva, T.G. Omelchenko, E.A. Zabirnyk, A.S. Markelova, E.V. Panibrattseva, S.G. Borovoy, I.A. Gubina-Vaculik, G.I. Oleynik, G.A. 2019-06-12T13:16:20Z 2019-06-12T13:16:20Z 2015 Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model / E.A. Shchegelskaya, T.G. Grigorieva, E.A. Omelchenko, A.S. Zabirnyk, E.V. Markelova, S.G. Panibrattseva, I.A. Borovoy, G.I. Gubina-Vaculik, G.A. Oleynik // Вiopolymers and Cell. — 2015. — Т. 31, № 5. — С. 387-394. — Бібліогр.: 13 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.0008FB https://nasplib.isofts.kiev.ua/handle/123456789/152613 576.08:576.524:57.089.6 Aim. To study migration of syngeneic bone marrow MSCs and skin fibroblasts (FB), labeled by fluorochromes, after intravenous (IV) and local transplantation in a rat burn wound model (BWM). Methods. Rats were divided into 3 groups: C – without burn + IV injection of labeled MSCs and FBs mixture; O1 – BWM + IV injection of labeled MSCs and FBs mixture; O2 – BWM + fibrin matrix filled with labeled cell mixture. MSCs were labeled by green fluorochrome, and fibroblasts – by red one. The presence of labeled cells in cryocuts of the skin, liver, kidney and bone marrow was assessed on the 3 and 7 days after transplantation. Results. Skin FBs selective migration to the regenerating burn wound and MSCs accumulation in the kidneys were found in rats of group O1 on day 7 after the IV injection. The labeled cells proliferated in the transplanted fibrin matrix and participated in the wound regeneration. MSCs partly migrated to the bone marrow after the IV injections. Conclusions. IV transplanted syngeneic bone marrow MSCs and skin FBs (passage 0) migrate to the burn wound and participate in the healing. Migration of bone marrow MSCs in the kidneys can prevent kidney failure after burn. Мета. Вивчення міграції сингенних МСК кісткового мозку і фібробластів (ФБ) шкіри, мічених флюорохромами, після внутрішньовенної (ВВ) та локальної трансплантації у щурів з моделлю післяопікової рани (МПР). Методи. Щури були розділені на 3 групи: К– без опіку + ВВ введення мічених МСК і ФБ; О1 – МПР + ВВ мічених МСК і ФБ; О2 – МПР + фібринова матриця з сумішшю мічених МСК і ФБ. МСК фарбували зеленим флюорохромом, а фібробласти – червоним. Розподіл мічених клітин на кріозрізах шкіри, печінки, нирок і кісткового мозку щурів вивчали на 3 і 7 добу після трансплантації. Результати. Виявлена вибіркова міграція ФБ шкіри в післяопікову рану і накопичення МСК КМ в нирках тварин групи О1 на 7 добу після ВВ введення мічених МСК і ФБ. У трансплантованій на рану фібриновій матриці мічені клітини проліферували та брали участь у процесі регенерації рани. При ВВ введенні МСК КМ спостерігалася їх міграція в кістковий мозок. Висновки. Системно введені сингенні МСК КМ і ФБ шкіри (0 пасаж) мігрують в зону післяопікової рани і беруть участь в її загоєнні. Міграція МСК КМ в нирки може попередити ниркову недостатність, що розвивається після опіку. Цель. Изучить миграции сингенных МСК костного мозга и фибробластов (ФБ) кожи, меченых флюорохромами, после их внутривенной (ВВ) и локальной трансплантации крысам с моделью послеожоговой раны (МПР). Методы. Крыс разделили на 3 группы: К – без ожога + ВВ введение смеси меченых МСК и ФБ; О1 – МПР + ВВ введение смеси меченых МСК и ФБ; О2 – МПР + фибриновая матрица с мечеными МСК и ФБ. МСК метили зеленым флюорохромом, фибробласты – красным. Наличие меченых клеток в коже, печени, почках и костном мозге крыс оценивали на 3 и 7 сутки после трансплантации на криосрезах и мазках. Результаты. Обнаружена избирательная миграция ФБ кожи в зону раны и накопление МСК КМ в почках крыс группы О1 на 7 сутки после ВВ введения меченых МСК и ФБ. В трансплантированной на рану фибриновой матрице меченые клетки активно делились и участвовали в ее регенерации. При ВВ введении МСК КМ частично мигрировали в костный мозг. Выводы. Системно трансплантированные сингенные МСК КМ и ФБ кожи 0 пассажа мигрируют в зону послеожоговой раны и участвуют в ее заживлении. Миграция МСК КМ в почки может предупредить вызванную ожогом почечную недостаточность. en Інститут молекулярної біології і генетики НАН України Вiopolymers and Cell Biomedicine Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model Міграція мічених МСК кісткового мозку і фібробластів шкіри після їх системної та місцевої трансплантації у щурів з моделлю післяопікової рани Миграция меченых МСК костного мозга и фибробластов кожи после их системной и местной трансплантации у крыс с моделью послеожоговой раны Article published earlier |
| spellingShingle | Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model Shchegelskaya, E.A. Grigorieva, T.G. Omelchenko, E.A. Zabirnyk, A.S. Markelova, E.V. Panibrattseva, S.G. Borovoy, I.A. Gubina-Vaculik, G.I. Oleynik, G.A. Biomedicine |
| title | Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model |
| title_alt | Міграція мічених МСК кісткового мозку і фібробластів шкіри після їх системної та місцевої трансплантації у щурів з моделлю післяопікової рани Миграция меченых МСК костного мозга и фибробластов кожи после их системной и местной трансплантации у крыс с моделью послеожоговой раны |
| title_full | Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model |
| title_fullStr | Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model |
| title_full_unstemmed | Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model |
| title_short | Migration of labeled bone marrow MSCs and skin fibroblasts after systemic and local transplantation in rat burn wound model |
| title_sort | migration of labeled bone marrow mscs and skin fibroblasts after systemic and local transplantation in rat burn wound model |
| topic | Biomedicine |
| topic_facet | Biomedicine |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/152613 |
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