TGF-β1 expression by glioma C6 cells in vitro
The aim of the work was to study the impact of fetal rat brain cell supernatant (FRBCS) on the expression of transforming growth factor β1 (TGF-β1) and p53 in C6 cells of rat glioma in vitro. Materials and Methods: FRBCS was obtained from suspensions of fetal rat brain cells on the 14th (E14) day of...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
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| Цитувати: | TGF-β1 expression by glioma C6 cells in vitro / L.D. Liubich, L.M. Kovalevska, M.I. Lisyany, V.M. Semenova, T.A. Malysheva, L.P. Stayno, V.V. Vaslovych // Experimental Oncology. — 2017 — Т. 39, № 4. — С. 258–263. — Бібліогр.: 40 назв. — англ. |
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irk-123456789-1385832018-06-20T03:07:53Z TGF-β1 expression by glioma C6 cells in vitro Liubich, L.D. Kovalevska, L.M. Lisyany, M.I. Semenova, V.M. Malysheva, T.A. Stayno, L.P. Vaslovych, V.V. Original contributions The aim of the work was to study the impact of fetal rat brain cell supernatant (FRBCS) on the expression of transforming growth factor β1 (TGF-β1) and p53 in C6 cells of rat glioma in vitro. Materials and Methods: FRBCS was obtained from suspensions of fetal rat brain cells on the 14th (E14) day of gestation. C6 glioma cells were cultured for 48 h in the presence of FRBCS or FRBCS + anti-TGF-β1 monoclonal antibody. Immunocytochemical staining for TGF-β1 and p53 was performed. Results: The proportion of TGF-β1-immunopositive tumor cells in C6 glioma cultures was statistically significantly higher than in the control cell cultures of normal fetal rat brain. FRBCS reduced the proportion of TGF-β1-immunopositive tumor cells and increased the proportion of p53-immunopositive cells in C6 glioma cultures. In cells cultured with FRBCS + anti-TGF-β1 monoclonal antibody, the above effects of FRBCS were abrogated. Conclusion: The obtained results suggest that TGF-β1 seems to be responsible for decrease in TGF-β1 expression and increase in p53 expression in C6 glioma cells treated with FRBCS. 2017 Article TGF-β1 expression by glioma C6 cells in vitro / L.D. Liubich, L.M. Kovalevska, M.I. Lisyany, V.M. Semenova, T.A. Malysheva, L.P. Stayno, V.V. Vaslovych // Experimental Oncology. — 2017 — Т. 39, № 4. — С. 258–263. — Бібліогр.: 40 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/138583 en Experimental Oncology Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine |
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English |
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Original contributions Original contributions |
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Original contributions Original contributions Liubich, L.D. Kovalevska, L.M. Lisyany, M.I. Semenova, V.M. Malysheva, T.A. Stayno, L.P. Vaslovych, V.V. TGF-β1 expression by glioma C6 cells in vitro Experimental Oncology |
| description |
The aim of the work was to study the impact of fetal rat brain cell supernatant (FRBCS) on the expression of transforming growth factor β1 (TGF-β1) and p53 in C6 cells of rat glioma in vitro. Materials and Methods: FRBCS was obtained from suspensions of fetal rat brain cells on the 14th (E14) day of gestation. C6 glioma cells were cultured for 48 h in the presence of FRBCS or FRBCS + anti-TGF-β1 monoclonal antibody. Immunocytochemical staining for TGF-β1 and p53 was performed. Results: The proportion of TGF-β1-immunopositive tumor cells in C6 glioma cultures was statistically significantly higher than in the control cell cultures of normal fetal rat brain. FRBCS reduced the proportion of TGF-β1-immunopositive tumor cells and increased the proportion of p53-immunopositive cells in C6 glioma cultures. In cells cultured with FRBCS + anti-TGF-β1 monoclonal antibody, the above effects of FRBCS were abrogated. Conclusion: The obtained results suggest that TGF-β1 seems to be responsible for decrease in TGF-β1 expression and increase in p53 expression in C6 glioma cells treated with FRBCS. |
| format |
Article |
| author |
Liubich, L.D. Kovalevska, L.M. Lisyany, M.I. Semenova, V.M. Malysheva, T.A. Stayno, L.P. Vaslovych, V.V. |
| author_facet |
Liubich, L.D. Kovalevska, L.M. Lisyany, M.I. Semenova, V.M. Malysheva, T.A. Stayno, L.P. Vaslovych, V.V. |
| author_sort |
Liubich, L.D. |
| title |
TGF-β1 expression by glioma C6 cells in vitro |
| title_short |
TGF-β1 expression by glioma C6 cells in vitro |
| title_full |
TGF-β1 expression by glioma C6 cells in vitro |
| title_fullStr |
TGF-β1 expression by glioma C6 cells in vitro |
| title_full_unstemmed |
TGF-β1 expression by glioma C6 cells in vitro |
| title_sort |
tgf-β1 expression by glioma c6 cells in vitro |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| publishDate |
2017 |
| topic_facet |
Original contributions |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/138583 |
| citation_txt |
TGF-β1 expression by glioma C6 cells in vitro / L.D. Liubich, L.M. Kovalevska, M.I. Lisyany, V.M. Semenova, T.A. Malysheva, L.P. Stayno, V.V. Vaslovych // Experimental Oncology. — 2017 — Т. 39, № 4. — С. 258–263. — Бібліогр.: 40 назв. — англ. |
| series |
Experimental Oncology |
| work_keys_str_mv |
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| first_indexed |
2025-07-10T06:06:52Z |
| last_indexed |
2025-07-10T06:06:52Z |
| _version_ |
1837238968039505920 |
| fulltext |
258 Experimental Oncology 39, 258–263, 2017 (December)
TGF-β1 EXPRESSION BY GLIOMA C6 CELLS IN VITRO
L.D. Liubich1, *, L.M. Kovalevska2, M.I. Lisyany1, V.M. Semenova1, T.A. Malysheva1, L.P. Stayno1,
V.V. Vaslovych1
1SI “Romodanov Neurosurgery Institute, National Academy of Medical Sciences of Ukraine”,
Kyiv 04050, Ukraine
2R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine,
Kyiv 03022, Ukraine
The aim of the work was to study the impact of fetal rat brain cell supernatant (FRBCS) on the expression of transforming growth
factor β1 (TGF-β1) and p53 in C6 cells of rat glioma in vitro. Materials and Methods: FRBCS was obtained from suspensions
of fetal rat brain cells on the 14th (E14) day of gestation. C6 glioma cells were cultured for 48 h in the presence of FRBCS or FRBCS
+ anti-TGF-β1 monoclonal antibody. Immunocytochemical staining for TGF-β1 and p53 was performed. Results: The proportion
of TGF-β1-immunopositive tumor cells in C6 glioma cultures was statistically significantly higher than in the control cell cultures
of normal fetal rat brain. FRBCS reduced the proportion of TGF-β1-immunopositive tumor cells and increased the proportion
of p53-immunopositive cells in C6 glioma cultures. In cells cultured with FRBCS + anti-TGF-β1 monoclonal antibody, the above
effects of FRBCS were abrogated. Conclusion: The obtained results suggest that TGF-β1 seems to be responsible for decrease
in TGF-β1 expression and increase in p53 expression in C6 glioma cells treated with FRBCS.
Key Words: transforming growth factor β1, p53, fetal rat brain cells supernatant, glioma C6, cell cultures, immunocytochemistry.
According to the National Cancer Registry, the
incidence of malignant brain tumors in Ukraine
in 2014 was 5.2 per 100,000 of population, and mor-
tality — 3.9 per 100,000 [1]. Among primary tumors
of central nervous system, glial tumors predominate.
The malignant glioblastomas are difficult to treat
because of their invasiveness and high recurrence
rate. In neurooncology, criteria for individualization
of patient treatment are actively developed by iden-
tifying a unique set of molecular changes in cancer
cells providing for personalized or targeted therapy.
As targets for personalized therapy of brain tumors,
the markers of proliferation, neuroepithelial stem cell
markers, regulators of cell proliferation, differentia-
tion, survival and/or apoptosis are considered [2].
The use of neural stem and progenitor cells (NSC/
NPC) is one of the approaches for the treatment of gli-
omas [3–6]. The similarities between NSC and brain
cancer stem cells (CSC) are being studied [7–13].
It is believed that NSC/NPC may be used to induce
long-term antitumor response by stimulating the im-
mune system and for delivery of various biological
agents to tumor [14, 15]. In the setting of experi-
mental glioblastomas, it is shown that the NPC can
migrate to the tumor inducing tumor cell death [15].
The use of NPC prolongs the survival of animals
or almost completely inhibits glioma growth [16], but
the mechanism of NPC antitumor effects remains
unclear.
The cultures of tumor cells are widely used for
developing new approaches to the treatment of brain
cancer [17]. In previous studies, we demonstrated
cytotoxic and antimitotic effects of fetal rat brain cell
supernatant (FRBCS) on 101.8 and C6 glioma cells
in vitro [18, 19] with reduction of the number of Ki-
67-immunopositive cells [20] and CD133-positive
cells in C6 glioma cell culture [21]. These proper-
ties seem to be mediated by soluble factors such
as TGF-β produced by NSC/NPC [18, 22, 23].
TGF-β is known as an important mediator of the ma-
lignant phenotype of human brain gliomas [24] while
TGF-β signaling is involved in the regulation of prolif-
eration, differentiation and survival/or apoptosis [25].
The antiproliferative effect of TGF-β1 on epithelial cells
in the early stages and promoting effect in the later
stages of tumor growth was shown [26]. It is believed
that TGF-β1 signaling is a potential target for antitu-
mor therapy, while the expression of TGF-β increases
significantly in gliomas with high degree of malignan-
cy [25, 27, 28]. Inhibitors of TGF-β1 signaling reduce
viability and invasive properties of gliomas modeled
in animals [25].
The study of the possible pro-apoptotic effects
of FRBCS and the expression of well-known tumor
suppressor protein p53 is no less important. P53 func-
tions as a transcription factor that regulates genes
involved in the cellular response to stress; activation
of p53 leads to the induction of DNA repair, cell cycle
arrest and apoptosis, whereas loss of p53 responses
due to mutations promotes uncontrolled cell prolifera-
tion [29].
The aim of the work was to study the impact
of FRBCS on the expression of TGF-β1 and p53 in gli-
oma cells in vitro.
MATERIALS AND METHODS
Cell lines. Rat glioma cells C6 were obtained
from the Bank of Cell Lines from Human and Animal
Tissues, R.E. Kavetsky Institute of Experimental
Submitted: November 18, 2016.
*Correspondence: E-mail: Lyubichld@gmail.com
Abbreviations used: CSC — cancer stem cells; FRBCS — fetal rat
brain cell supernatant; NPC — neural progenitor cells; NSC — neu-
ral stem cells; TGF-β1 — transforming growth factor β1.
Exp Oncol 2017
39, 4, 258–263
ORIGINAL CONTRIBUTIONS
Experimental Oncology 39, 258–263, 2017 (December) 259
Pathology, Oncology and Radiobiology, the National
Academy of Sciences, Kyiv). The cells of fetal rat brain
(E14) were obtained under the protocol [21]. The vi-
ability of cells was determined in a standard test with
0.2% trypan blue (Merck, Germany) [21]. 1•106 cells
were applied onto the adhesive coverslips coated
with polyethylenimine (Sigma, USA), which were
placed in Petri dishes and cultured in DMEM medium
(2 ml, Sigma, USA), supplemented with 1% fetal calf
serum (Sigma, USA), glucose (400 mg%) and insulin
(0.2 U/ml). Cells were cultured in a CO2-incubator
(37 °С, 95% humidity and 5% CO2) and observed
in inverted microscope (Eclips TS 100, Japan) with
microphotographic registration.
FRBCS. Fetal rat brain cells removed on the 14th
(E14) day of gestation were cultured as previously
described [18] and supernatant (0.10 mg/ml) was col-
lected. Earlier we showed that 49–50% of E14 cells are
nestin-immunopositive [30] and 37–40% — CD133-
immunopositive [21], i.e. these cells are positive for
NSC markers.
Monoclonal antibodies. Monoclonal antibody
to TGF-β1 (anti-transforming growth factor-β1,
clone 9016.2; Sigma, USА) and rabbit antibody
to р53 (anti-TP53 antibody; Sigma- Aldrich, USА)
were used. For FRBCS neutralization, FRBCS
(0.10 mg/ml) was mixed with monoclonal anti-
body to TGF-β1 (0.10 µg/ml) and incubated for
20–30 min prior to addition to the experimental
cultures.
Immunocytochemical staining for TGF-β1 and
p53. Cells fixed on coverslips were rehydrated, incu-
bated in 0.1% solution of Triton X-100 (Sigma, USA)
at room temperature for 30 min, and then washed
three times for 5 min in 0.01 M phosphate buffer
(рН 6.0). To block endogenous peroxidase, the cov-
erslips were incubated in the dark for 10 min with 3%
H2O2 solution and then rinsed for 5 min in phosphate
buffer. To block nonspecific background staining,
coverslips were incubated 5 min with 1% solution
of bovine serum albumin (Sigma, USА). Mouse
monoclonal antibody to TGF-β1 or rabbit antibody
to p53 were applied at a dilution of 1:100 for 60 min
at room temperature. After triple washing in buffer,
the secondary antibody (goat antimouse/antirab-
bit IgG peroxidase conjugated (Dako, Denmark)
at a dilution of 1:200 were applied for 30 min at room
temperature followed by treatment with diaminoben-
zidine solution (Dako, Denmark) for 2 min and the
development of specific coloration was controlled
under the microscope. After washing with distilled
water, the cells were stained with hematoxylin and
the specimens were embedded in balsam. Parallel
studies were performed with positive and negative
controls. The stained specimens were examined
under AxioImager A2 microscope (Carl Zeiss Mi-
croscopy GmbH, Germany) with a broadband filter
equipped with camera AxioCam MRc5 (Carl Zeiss
Microscopy GmbH, Germany). TGF-β1- and p53-
immunopositive and negative cells were counted
Fig. 1. Change in the TGF-β1 expression in C6 glioma cells treated with FRBCS. Immunocytochemical staining for TGF-β1 and coun-
terstaining with hematoxylin, × 2000, immersion: a — C6 cells, control; b — culture of fetal rat brain cells (E14), control; c — C6 cells
incubated with FRBCS (E14), 0.10 mg/ml, 48 h; d — C6 cells incubated with a mixture of FRBCS (E14, 0.10 mg/ml) and antibody
to TGF-β1 (0.10 mg/ml), 48 h
260 Experimental Oncology 39, 258–263, 2017 (December)
in 10 representative fields of view with standard
measurement scale (object-micrometer). Digital
images analysis was performed using the software
“Zen Lite 2012” (Germany).
Statistical analysis. Statistica 8.0, software
StatSoft, Inc. (2007) was used for nonparametric
Mann — Whitney U-test for comparison of indepen-
dent groups. Normality of data distribution was deter-
mined Shapiro — Wilkie test. A statistically significant
difference was considered when p < 0.05, statistically
highly significant — when p < 0.01.
RESULTS AND DISCUSSION
Immunostaining for TGF-β1 revealed 48.81 ± 7.91%
of TGF-β1-immunopositive cells in control glioma
C6 specimens (Fig. 1, a; Fig. 2) that exceeds signifi-
cantly the percentage of TGF-β1-immunopositive cells
in, control cell cultures of normal fetal rat brain (E14)
(22.04 ± 2.33%, Mann — Whitney U-test, p = 0.048;
Fig. 1, b; Fig. 2).
After 48 h of incubation of C6 glioma cultures with
FRBCS in a concentration of 0.10 mg/ml, the fraction
of TGF-β1-immunopositive tumor cells statistically
significantly decreased to 29.18 ± 7.24% (Mann —
Whitney U-test, p = 0.0003 compared to control;
Fig. 1, c; Fig. 2). The incubation of C6 cells with
a mixture of FRBCS and monoclonal antibodies
to TGF-β1 abrogated the decrease of TGF-β1-
immunopositive fraction in cultures treated with
FRBCS as a single agent (p = 0.05, Mann — Whitney
U-test; Fig. 1, d; Fig. 2). Earlier we reported that
FRBCS contains TGF-β1 in an amount of 12.0 pg/
ml [18]. Therefore, the decrease of TGF-β1-
immunopositive cell count in glioma C6 culture
after incubation with FRBCS may be presumably
explained by the effects of TGF-β1 as the compo-
nent of FRBCS.
The amount of TGF-β1-immunopositive cells, %
%
10
20
30
40
50
60
70
80
1 2
p = 0.048
p = 0.005
p = 0.0003
p = 0.007
Groups
M
M ± m
M ± d
3 4
Fig. 2. Quantitative indicators of changes in the expression
of TGF-β1 in C6 glioma cells treated with FRBCS, 0.10 mg/ml,
48 h: 1 — glioma C6 cells, control; 2 — glioma C6 cells incu-
bated with FRBCS (E14), 0.10 mg/ml, 48 h; 3 — glioma C6 cells
incubated with a mixture of FRBCS (E14, 0.10 mg/ml) and
antibody to TGF-β1 (0.10 mg/ml), 48 h; 4 — culture of fetal rat
brain cells (E14), control. M — mean value; m — standard error
of the mean; d — standard deviation from the mean
We also examined the expression of p53 in C6 gli-
oma cells treated with FRBCS. In control C6 cells
the fraction of p53-immunopositive cells was
8.94 ± 6.05% (Fig. 3, a; Fig. 4). After 48 h of incuba-
tion with FRBCS in a concentration of 0.10 mg/ml the
fraction of p53-immunopositive C6 cells statistically
significantly increased to 18.47 ± 10.62% (Mann —
Whitney U-test, p = 0.01 compared to control;
Fig. 3, b; Fig. 4). Again, this effect of FRBCS
was сanceled when monoclonal ant ibodies
to TGF-β1 were added (p = 0.003 compared with
cultures incubated only with FRBCS, Mann — Whit-
ney U-test; Fig. 3, c; Fig. 4). That is, while adding
to the C6 glioma cells a mixture of FRBCS and
monoclonal antibody to TGF-β1 the neutralization
Fig. 3. Change of the p53 expression in C6 glioma cells treated
with FRBCS. Immunocytochemical staining for p53 and coun-
terstaining with hematoxylin, × 2000, immersion: a — C6 cells,
control; b — C6 cells incubated with FRBCS (E14), 0.10 mg/ml,
48 h; c — C6 cells incubated with a mixture of FRBCS (E14,
0.10 mg/ml) and antibody to TGF-β1 (0.10 mg/ml), 48 h
Experimental Oncology 39, 258–263, 2017 (December) 261
of FRBCS effect took place because of binding
molecules of TGF-β1 with specific monoclonal an-
tibodies. In our view, this indicates that the estab-
lished effect (increase of p53-immunopositive cells
in glioma C6 cultures) after incubation with FRBCS
takes place due to the influence of TGF-β1, which
is a component of FRBCS.
The amount of p53-immunopositive cells
%
-5
0
5
10
15
20
25
30
35
p = 0.01 p = 0.003
M
M ± m
M ± d
1 2
Groups
3
Fig. 4. Quantitative indicators of changes in the expression
of p53 in C6 glioma cells treated with FRBCS, 0.10 mg/ml,
48 h): 1 — glioma C6 cells, control; 2 — glioma C6 cells incubated
with FRBCS (E14), 0.10 mg/ml, 48 h; 3 — glioma C6 cells incu-
bated with a mixture of FRBCS (E14, 0.10 mg/ml) and antibody
to p53 (0.10 mg/ml), 48 h; M — mean value; m — standard error
of the mean; d — standard deviation from the mean
So we have shown, firstly: two-fold increase
of cells expressing TGF-β1, in C6 glioma cell cultures
compared with cell cultures of normal fetal brain;
secondly: reduction of TGF-β1-immunopositive
cell number in C6 glioma cultures after FRBCS
treatment. The obtained results are generally
agreed with the known data about mutations of the
components of TGF-β1-signaling pathways in cells
of gliomas, as well as autocrine-paracrine mecha-
nism of TGF-β1, effects [24, 31].
In this regard it should be noted that under physi-
ological conditions TGF-β plays an important role
in the embryo- and morphogenesis and in main-
tenance of tissue homeostasis [31, 32]. In par-
ticular, according to immunohistochemical studies
of Pelton et al. [31], few TGF-β1-immunoreactive
cells, and a significant number of TGF-β2-, TGF-β3-
immunoreactive cells were identified in embryonic
tissue of the central nervous system of mouse in fe-
tuses aged 12.5–18.5 days. The authors concluded
that the isoforms of the growth factor TGF-β during
embryogenesis of mammals work by both paracrine
and autocrine mechanism, regulate differentiation
(by stimulation or inhibition, depending on the type
of cells), stimulate the formation of extracellular
matrix, act as chemoattractant for certain cells,
and also induce mesoderm formation during early
development [31]. Other researchers have shown
that multipotent human, rat and mouse NPC are able
to produce all isotypes of TGF-β, in particular, TGF-β1,
TGF-β2, which may explain the immunosuppressive
nature of these cells [22, 23].
In setting of pathology, TGF-β has a dual role.
On the one hand, TGF-β is an important mediator
of the malignant phenotype of human brain gliomas
being involved in the regulation of proliferation, differ-
entiation and survival/or apoptosis [25], modulation
of invasiveness, angiogenesis, evasion of immune
control and maintenance of CSC in brain [24, 33–35].
On the other hand, TGF-β is a strong inhibitor of pro-
liferation of epithelial cells, astrocytes, immune cells
and is considered as tumor-suppressive factor [24,
36]. Other authors reported TGF-β1 antiprolifera-
tive effect of TGF-β1 in a number of epithelial cells
in the early stages and promoter effect in the later
stages of tumor growth [26, 32]. Tumor-suppressive
function of TGF-β includes inhibition of proliferation,
induction of apoptosis, regulation of autophagy, but
with the development of tumors the response of cells
to TGF-β shifts. As a result TGF-β acts as a potential
promoter of cell motility, invasion, metastasis and
CSC maintenance in brain [37]. In carcinogenesis,
TGF-β SMAD-dependent signaling pathway cor-
relates with antiproliferative and tumor-suppressive
functions of TGF-β, while SMAD-independent way
is involved in pro-tumor functions of TGF-β [26, 32].
This occurs as a result of mutations of canonical TGF-
β-signaling pathway elements in malignant gliomas,
which allow avoiding the antiproliferative influence
of TGF-β, thereby contributing to its promoting
activity [24]. TGF-β modulates the response of glio-
blastoma cells by autocrine way, all three isoforms
of TGF-β are expressed being biologically active
in glioblastoma cells [24]. The total result of this multi-
directional (antiproliferative or stimulating) effect
of the TGF-β can vary between different specimens
of tumors and even between different parts of the
same tumor [24].
It is believed that TGF-β-signaling is a potential
target for the therapy of cancer as the expression
of TGF-β isoforms increases significantly in gliomas
with high degree of malignancy [24, 25, 27, 28, 32,
38], helping tumor to avoid immune recognition via
various mechanisms, including inhibition of CD8+ cy-
totoxic lymphocytes and natural killer cells [39] and
stimulation of the generation of T regulatory cells.
The three levels of therapeutic strategy of TGF-β-
signaling inhibition are considered: the ligand itself,
the ligand — receptor interactions and the intracel-
lular signaling cascade [32]. Inhibitors of TGF-β-
signaling, namely, neutralizing monoclonal antibodies
to TGF-β used in combination with vaccine based
on glioma-associated antigen peptides reduce viability
and invasive properties of gliomas in animals [25] con-
tributing to lengthening the average life span mice with
GL261 glioma [27]. The knockdown of TGF-β receptor
type 2 reduced invasiveness of glioma CSC in vivo [40].
We have demonstrated that FRBCS reduced the
proportion of TGF-β1-immunopositive C6 glioma
262 Experimental Oncology 39, 258–263, 2017 (December)
cells and increased the proportion of p53-immu-
nopositive cells. Nevertheless, a mixture of FRBCS
and anti-TGF-β1 monoclonal antibody neutralized
the above biological effects of FRBCS in C6 glioma
cells. Thus, our data suggest that TGF-β1 seems
to be responsible for decrease in TGF-β1 expression
and increase in p53 expression in C6 glioma cells
treated with FRBCS.
The obtained results, in our opinion, may become
the basis for further research for the purpose of theo-
retical substantiation of complex pathogenetic therapy
of the patients with gliomas including the use of the
products derived from fetal neurogenic cells.
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