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2D- and 3D-cell culture

Article  in  Biopolymers and Cell · January 2011

DOI: 10.7124/bc.00007D

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ISSN 0233–7657. Biopolymers and Cell. 2011. Vol. 27. N 1. P. 17–24 (Translated from Russian) doi:10.7124/bc.00007D

2D- and 3D-cell culture

A. I. Khoruzhenko

Institute of Molecular Biology and Genetics NAS of Ukraine

150, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03680
a.i.khoruzhenko@imbg.org.ua

The cultivation of mammalian cells in three-dimensional conditions acquires a priority in a variety of

biomedical applications. In the areas of toxicology and anticancer drug development it concerns a
significant difference of responses to proapoptotic factors of the cells cultured in 2D versus 3D
environment. Besides, the clear-cut differences have been found in cell polarity, cytoskeleton structure,
distribution of receptors to wide range of hormones, growth factors, etc. in mammalian cells depending on
culture conditions. It is resulted in different response of cultured cells to extracellular stimuli. Multicellular
spheroids are regarded presently as the most convenient model of solid tumour growth in vitro. The
cultivation of thyroid follicles, mammary acini and other structure units, maintaining initial tissue
organization, allows studying the behavior, biochemical features and gene profile of differentiated cells. On
the other hand, 3D cultures have some limitations in comparison with a well established monolayer culture.
The advantages and disadvantages of each type of cultures and their application in biological and medical
researches will be discussed in this review.
Key words: 2D and 3D cultivation, resistance, differentiation.

Brief overview of creating the method of cell cultures were obtained by placing the fragment of
culture. The method of cell culture is a powerful tissue into the plasma with subsequent formation of a
instrument of medical and biological studies. The first fibrin clot or into the agar. This cultivation lasted at
fundamental articles, devoted to the elaboration of this least 20 days [4]. The decisive importance was attri-
approach, were written as far as 100 years ago. The buted to the fact that it was possible to isolate cell
selection of cultivation conditions, which are standard cultures not only from different tissues of the organism,
nowadays (temperature, humid gas phase, aseptics but also from primary cultivated cells due to their pas-
technology, standardized cultural nutrient media), saging, as shown on secondary and tertiary cultures of
required significant efforts, which was reflected in the thyroid gland, spleen, sarcoma cells [4].
works of Lambert, Carrel, and others [1–4]. The very It was practically at the same time that several labo-
possibility of existence of living cells outside the initial ratories discovered the ability of cells to have proli-
organism was first shown by Loeb in 1897, at the time feration in the culture conditions. Another interesting
of the first successful attempt to transplant the skin fact was revealed in the observation that the cells of the
fragments of a guinea pig embryo into agar, on nervous system of the frog embryo were capable of
coagulated serum and into the organism of an adult differentiating ex vivo, namely, they formed long ner-
animal [2]. With time the source of obtaining cultures vous endings [4]. Besides, it was established in 1912
was not only the embryonic tissue, as cell cultures were that cultivated mammalian cells; in particular, the cells
isolated from different tumors as well [3]. The first of lymph nodes and bone marrow of a guinea pig are
capable of producing antibodies [5].
Ó Institute of Molecular Biology and Genetics NAS of Ukraine, 2011

17
KHORUZHENKO A. I.

The next stage of elaborating the method of cell is co-cultivation of epidermal keratinocytes and dermal
culture was the attempt of isolating pure cell cultures of fibroblasts) [11].
a certain type. Thus, primary cultures, consisting either The sources of obtaining cell cultures. The cell
of amoeboid cells or round cells [6], were isolated from cultures of mammals, including humans, are most
the stroma of heart of the chicken embryo. Maksimov, widely used in the investigations in the biomedical
who formulated the unitary theory of hematopoiesis sphere. Therefore, the main attention of this review will
and was the first to use the term “a stem cell”, be paid to the abovementioned cultures. They are
investigated human connective tissue by the method of obtained both from the normal tissue and patholo-
cell culture [7]. gically modified organs and tissues, including benign,
Further application of the cellular material of malignant tumors, tissues with inflammation, disorder
chicken embryo allowed isolating primary cultures of of metabolic processes, etc.
different types of cells, while the application of cell Primary cultures of cells, obtained from the tissue
cultures of rodents facilitated the application of esta- of the organism, have a number of advantages: firstly,
blished cellular lines [8]. The possibility of supporting this is direct origin from the cells of the primary orga-
established lines, originating from human tumor cells, nism and relative simplicity of obtaining two-dimen-
was shown on the example of HeLa cells [9]. The sional primary cultures. The cultivation of mammalian
classic studies of Hayflik revealed temporal limits of cells in three-dimensional conditions while preserving
life of normal chicken embryo cells in the culture [10]. initial tissue organization allows maximal approxi-
Further rapid development of the method of cell culture mation of experiment conditions to those of the orga-
demonstrated its suitability not only for biomedicine, nism. In medicine primary cultures are used to deter-
but also for studies in the sphere of pharmacology, mine the karyotype and in some other investigations
ecology, and veterinary science. [13]. On the other hand, there are some limits of their
Terminology. Prior to laying out the method of application, including the need of the source to obtain
cultivation, let us present some most frequent terms cultures, a lower level of reproducing organisms; limi-
according to Freshney’s manual [11]. A tissue culture is ted number of possible reduplication of the population
a generic term that refers to both organ culture and cell for cells from the normal tissue. These problems may
culture. An organ culture is always meant to be the be partially solved by using cell lines, obtained from
three-dimensional culture of disaggregated tissue, pre- normal, benign or malignized tumor tissues, for the
serving some or all the histological specificities of the study in the mechanisms of malignant transformation
primary tissue; the cell culture is the culture, initiated of cells, trial of medical preparations, in particular,
from dispersed cells, isolated from the primary tissue, suppressors of tumor growth, revealing differences in
primary cultures or cell lines due to enzymatic, che- physiological and biochemical indices of tumor and
mical or mechanical disaggregation. According to normal cells. The cells of lines are cultivated both in
Adams’s manual [12] the cell culture should be con- two-dimensional and three-dimensional conditions
sidered as primary one, when it is isolated directly from [14]. The known cell lines are widely used in research
the organism; and its further cultivation allows work, they are remarkable for a higher level of repro-
obtaining cell lines, which are generally heterogeneous ducibility in experiments compared to primary lines. It
in their composition. Cloning, physical or other its turn, the application of cell lines has some limits.
selection of cells from the cell line allows obtaining cell The phenotype and genotype of cultivated cells may
strains with specific properties [11, 12]. Histotypic differ considerably from those of the cells of the initial
cultures are the result of disaggregation of cellular tissue. In particular, the populations of malignant tu-
material for restoration of three-dimensional tissue-like mour cells are heterogeneous in their composition; the-
structure. Organotypic cultures are similar to refore, during cultivation there is selection of cells
histotypic ones, but their isolation requires the according to their capability to survive. Besides, the
application of several types of cells (a classic example work with several cell lines is accompanied with the
problem of their cross-contamination. The investi-

18
 2D- AND 3D-CULTURE OF CELL

gations, performed by a number of authors, demonstra- Some authors used the cells of thyroid gland as an
ted that only 23 among 40 most widely used cell lines example to demonstrate that the loss of follicular orga-
of thyroid gland had a unique genetic profile. Some nization in conditions of the monolayer culture results
lines had common origin, while others were cross- in the loss of polarization of thyrocytes, considerable
contaminated by cells of other lines [15]. Cell lines are decrease in the ability to capture and transport iodine,
inapplicable for determination of individual sensitivity disorder of intercellular contacts and connection to the
to medical preparations (chemotherapeutic, first of all). basal membrane [19, 20]. Similar modifications were
The approaches to cultivation of cells of humans observed for the cells of other organs and tissues,
and mammals, their disadvantages and advantages. including mammary gland, kidneys, etc. [21, 22]. The
Organ cultures. The cultivation of small parts of tissue cultivation in three-dimensional conditions was sugge-
by the method of organ culture allows preserving the sted to solve these issues. The cultures, in which cells
initial histological structure which provides consi- are stimulated by relevant factor to form initial histo-
derable correlation with in vivo conditions. However, like structures, may be considered as an intermediate
after several days of cultivation the process of necrosis variant [23, 24].
is launched in the central zone of the explant, which is Besides, the use of collagen or gelatin substrate for
conditioned by insufficient access of oxygen and nut- floating monolayer cultures allows preserving the
rients. With time this process spreads to periphery as polarization of cultivated epithelial cells [25]. For
well. Special means are elaborated to overcome this instance, the work [25] presents the data on asymmetry
problem, ACUSYST-S chamber being among them of localization of receptors to the thyrotropic hormone.
[16]. The second approach presupposes the application The introduction of the latter to the cells, cultivated on
of miniorganic cultures. Precise microscopic equip- collagen gel, attached to the surface of plastic, did not
ment allows decreasing the size of grained fragment of result in any observed effect on the cells. If the gel with
thyroid tissue to 0.5 – 0.9 mm, besides, the content of cells is unfastened from the cultural surface, thyrocytes
oxygen is increased to 50 % which promotes rapid become sensitive to the effect of thyrotropic hormone.
reduction of the necrosis zone. Thus, the cultures of the Further elaboration of this approach led to the
normal tissue and malignant tumor of thyroid gland application of bicameral chambers, divided by the
were obtained [17]. membrane – transwells. The example of cell cultures,
Monolayer cultures and cultures on floating obtained from normal and tumor tissues of different
substrates. The method of monolayer cultivation is wi- organs, demonstrates that cells preserve their polarity
dely used in biomedical studies, in particular, with the during their cultivation on transwells [26, 27].
purpose of determining the mitotic potential of cells, Multicellular spheroids. Multicellular spheroids
change of their functional activity under the impact of are often used to simulate tumor growth, especially in
specific factors, for comparison of physiological and case of poorly differentiated tumors, further they are
biochemical characteristics of cells, obtained from nor- analyzed by biochemical, histological and immuno-
mal and tumor tissues. Monolayer cultures of dermal histochemical methods [28]. To obtain spheroids, the
fibroblasts are used for the analysis of human karyo- suspension of cells is placed into conditions, preven-
type for diagnostic purposes along with the suspension ting the adhesion of cells to growth surface. An agarose
culture of lymphocytes. However, this approach has a layer or siliconized surface is most frequently used for
significant drawback. During the cultivation in two- this purpose. Similar to organ cultures, the limitation of
dimensional conditions the investigated cells lose their this method of cultivation is insufficient access of
histological organization, which is finally reflected in oxygen and nutrients into central parts of the spheroid
their behavior and biochemical specificities. Besides, after it gets considerable size. Spheroid cultures are
the polarization of cultivated cells is violated. First of applied to analyze extracellular matrix due the
all, it is true regarding epithelial cells, which have similarity of spheroid cultures and solid tumors [30,
clearly differentiated functions of apical and basal 31]. Moreover, the application of spheroid cultures
membranes in vivo [18]. allowed revealing the difference in the interaction of

19
KHORUZHENKO A. I.

papillary and follicular thyroid carcinoma cells with lated to maintaining functional activity of the investi-
endothelial cells during the simulation of extravasation gated cells. On the other hand, this methodological
of tumor cells from the blood flow [32]. approach allows performing histological and immuno-
Cultivation in soft agar. Tumor potential of histochemical analysis of cultures regarding the pre-
cultivated cells is often revealed by their capability to sence and localization of certain antigens in the very
form colonies in soft agar. The essence of this test is the cells and in the extracellular matrix. Its limitation is the
determination of the number of cells, capable of giving alleged obtaining of sufficient amount of material from
independent beginning to a new colony which is dee- the tissues of the organism to initiate primary cultures.
med to be the model of initial stages of metastases in Similar to multicellular spheroids, the acquisition of
the conditions of the organism. Then the suspension of considerable sizes delays the access of oxygen and
investigated cells is placed into soft agar (the concen- nutrients into the central sites of the aggregate. The
tration of agarose is about 0.3 %) to secure independent maintaining of cells in the differentiated state (like for
growth of each cell. The efficiency coefficient of colo- other types of cultures) requires additional growth
ny-formation is calculated after certain time of culti- factors and hormones depending on the origin of
vation; for this purpose the ratio of the number of for- cultivated cells. Besides, during enzymatic disaggre-
med colonies and the number of introduced cells is gation of the tissue some structures are destroyed and
expressed in percentage terms [33]. Regardless of the unstructured parts may appear in the composition of
fact that the issue of the origin of metastases from one floating aggregates. However, taking into conside-
cell or from the colony of cells is yet to be discussed, ration the abovementioned issues this approach allows
this test is widely used in pharmacological studies of studying the properties of cells in conditions, maxi-
tumor suppressors [34]. Many authors demonstrated mally approximated to in vivo state. Thus, we have
that the sensitivity of tumor cells, cultivated in soft shown that the cultivation of human and murine
agar, to proapoptotic agents has better correlation with thyrocytes in conditions of monolayer culture leads to
the results, obtained in vivo, than the reaction of cells, significant content of thyroglobulin in the cells, which
cultivated in the monolayer. is the main index of their functional activity. The im-
Aggregates of follicles, acini, etc. In addition to the munochemical analysis revealed that in 10 days of
abovementioned, there is another approach to the cultivation in the monolayer culture only 30 % of cells
cultivation of mammalian cells in three-dimensional contain thyroglobulin, similar results were obtained
conditions. It presupposes the application of “func- during the investigation of cells, cultivated in the form
tional units” (follicles, acini, etc.) for the initiation of of multicellular spheroids. Contrary to them, all the
primary cultures with further preservation of the initial cells in the composition of follicle-like structures of
histological structure. For this purpose the follicles or floating aggregates of follicles demonstrate expressed
acini are “enucleated” from the initial tissue using positive immunohistochemical reaction to thyroglo-
matrix-degrading enzymes and they are either placed bulin. Moreover, the cells from the sites, which do not
on the non-adhesive substrate, or immersed into col- preserve follicle-like organization, did not contain
lagen gel (matrigel, etc.). It is possible to work with the thyroglobulin. Therefore, our studies revealed that
suspension of thyroid follicles in conditions of the cultivation of thyrocytes in the form of aggregates of
rotational culture [35]. However, if they are placed on follicles is closer to the conditions of the living
the layer of agarose or siliconized plastic, it is possible organism than cultivation in the form of a monolayer or
to achieve the formation of floating aggregates, preser- multicellular spheroids [36, 37].
ving the initial histological structure in many aspects. The application of two- and three-dimensional
These cultures have significant advantages from the cultures in biomedical studies. The problem of
standpoint of both cell physiology and methodological maintaining the differentiation of cells in the
approaches to the application of cultures in experi- monolayer culture. The histological structure of tissues
ments. In the majority of cases the preservation of the of the organism is closely related to the fulfillment of
initial structure of the tissue in culture conditions is re- specific functions of the cells. The structure of intes-

20
 2D- AND 3D-CULTURE OF CELL

tines secures the processes of resorption of nutrients, mensional cultures. During the incubation of cells, gro-
alveoli are responsible for gas exchange, muscle tissue wing in the three-dimensional culture, with antibodies
– for contractions, etc. The disorders in the functions of to beta 1-integrin, breast carcinoma cells acquired
organs are very often accompanied by changes in the non-malignized phenotype, lost the atypical form and
tissue organization. For instance, the substitution of the changed the type of growth. However, this effect was
muscle tissue with connective tissue fibrils complicates not reproduced in the monolayer culture [43].
muscle contractions, squamous metaplasia of thyro- On the next stage this group revealed that during
cytes impairs the function of the thyroid gland, the the three-dimensional cultivation the addition of
occurrence of atherosclerosis plaques impairs blood antibodies to beta 1-integrin hinders the transmission
circulation, etc. Therefore, the simulation of different of the intracellular signal from receptors of epidermal
processes in ex vivo conditions requires the consi- growth factor (EGF). On the other hand, the antibodies
deration of specificities of the initial tissue. to receptor EGF inhibit the activity of beta 1-integrin.
The application of monolayer cultures in the study This reciprocal effect was not achieved in the con-
of keratinocytes of skin of humans and mammals pro- ditions of two-dimensional culture [44]. The receptors
ved to be successful. It was possible not only to obtain to growth factors play a key role in the development of
reproducible cultures of cells, but also the impact of malignant tumors, and this is not the only conclusion of
certain factors (change in the concentration of calcium, the investigation of carcinogenesis mechanisms,
cultivation on the interface of liquid and gas phases, obtained using three-dimensional cultures. Usually
etc.) allows achieving the level of stratification of during metastasis the cells produce enzymes, splitting
layers of keratinocytes, corresponding to that in vivo. the fibrils of extracellular matrix [45]. The inhibitors of
Practically, the approach to obtaining of artificial ana- these enzymes are sometimes used in the clinical
logues of skin (including the application of cultivated practice. However, the model of three-dimensional
keratinocytes and fibroblasts) is based on the combi- cultures was used to demonstrate that even complete
nation of methods of two- and three-dimensional culti- inhibition of matrix metalloproteinases, serine protea-
vation, which is successfully applied in many labora- ses, cathepsin and other proteases do not prevent the
tories [38]. It was shown that in conditions of a fibrosarcoma cells from migrating. Their protruded
monolayer culture the endothelial cells contain von form becomes more round and due to amoeboid
Willebrand factor, which is one of the markers of their movement the cells keep “pushing” through the matrix
differentiation, and are capable of secreting the extra- fibrils. The behavior of these cells is the same after the
cellular matrix, partially reminding their matrix in vivo subcutaneous injection to mice [46]. Moreover, Mar-
[39]. On the other hand, these conditions promote the shall et alt. demonstrated that during blocking of
impairment of the capability of smooth muscles to con- Rho-signaling pathway the cells lose their capability of
tract [40]. As shown above, the content of thyro- amoeboid movement, independent from proteolytic
globulin in thyrocytes decreases sharply. Although in enzymes. Thus, the three-dimensional model of
some cases the accumulations of thyrocytes, formed cultivation was used to demonstrate the pathway of
under the impact of thyrotropic hormone in monolayer combined inhibition of metastasis of tumor cells [47]. It
culture, were considered to be follicle-like structures was revealed using fibroblasts that contrary to
[41]. monolayer culture these cells grow faster in the three-
The sensitivity of cells to proapoptotic factors in dimensional culture and become asymmetrical, similar
conditions of two and three-dimensional cultures. to in vivo conditions [48].
Recent trends of medical and biological researches Besides, the studies demonstrated the resistance of
testify to the advantages of the cultivation of cells in mesothelioma cells, cultivated in three-dimensional
three-dimensional conditions [42]. In 1997 Bissell’s conditions, to TRAIL (TNF – related apoptosis- indu-
group received a number of well-founded data on the cing ligand) in the complex with gemcitabine, while in
difference in the behavior of human breast carcinoma the monolayer culture the cells showed their resistance
cells, cultivated in the form of two and three-di- to these agents [49, 50]. Similar effect was observed

21
KHORUZHENKO A. I.

during the investigation of the impact of the complex of öèíñêèõ ïðèëîæåíèÿõ. Â îáëàñòè òîêñèêîëîãèè è ðàçðàáîòêè
ïðîòèâîîïóõîëåâûõ ïðåïàðàòîâ ýòî ñâÿçàíî ñî çíà÷èòåëüíîé
Bortezomib and TRAIL as proapoptotic agents on lung ðàçíèöåé îòâåòà êëåòîê, êóëüòèâèðóåìûõ â äâóõ- èëè òðåõìåð-
carcinoma cells A549. After the introduction of these íûõ óñëîâèÿõ, íà ïðîàïîïòîòè÷íûå ôàêòîðû. Êðîìå òîãî, âû-
ÿâëåíû ñóùåñòâåííûå îòëè÷èÿ â ïîëÿðèçàöèè êëåòîê, ñòðóê-
cells into the conditions of the monolayer culture the
òóðå öèòîñêåëåòà, ðàñïðåäåëåíèè ðåöåïòîðîâ ê øèðîêîìó
mentioned agents caused apoptosis, while in spheroid ñïåêòðó ãîðìîíîâ, ðîñòîâûõ ôàêòîðîâ è ò. ä. â êëåòêàõ ìëåêî-
cultures the investigated cells were resistant to these ïèòàþùèõ â çàâèñèìîñòè îò óñëîâèé êóëüòèâèðîâàíèÿ. Â êî-
íå÷íîì èòîãå, ýòî ïðîÿâëÿåòñÿ â ðàçëè÷íîé ðåàêöèè êóëü-
factors, which correlates with the results, obtained on òèâèðóåìûõ êëåòîê íà âíåêëåòî÷íûå ñòèìóëû. Íàèáîëåå àäåê-
laboratory animals [51]. âàòíîé ìîäåëüþ ðîñòà ñîëèäíûõ îïóõîëåé in vitro ñåé÷àñ ïðè-
Conclusion. Summing up the abovementioned, it íÿòî ñ÷èòàòü ìíîãîêëåòî÷íûå ñôåðîèäû. Êóëüòèâèðîâàíèå
ôîëëèêóëîâ ùèòîâèäíîé æåëåçû, àöèíóñîâ ìîëî÷íîé æåëåçû è
should be mentioned that each approach to the culti- äðóãèõ ñòðóêòóðíî-ôóíêöèîíàëüíûõ åäèíèö, ñîõðàíÿþùèõ èñ-
vation has both advantages and disadvantages. Howe- õîäíóþ îðãàíèçàöèþ òêàíè, ïîçâîëÿåò èçó÷àòü ïîâåäåíèå, áèî-
ver, the focus of selecting the conditions of the cul- õèìè÷åñêèå îñîáåííîñòè è ãåíåòè÷åñêèé ïðîôèëü äèôôå-
ðåíöèðîâàííûõ êëåòîê. Ñ äðóãîé ñòîðîíû, òðåõìåðíûå êóëü-
tivation should be placed at the initial histological or- òóðû èìåþò ðÿä îãðàíè÷åíèé ïî ñðàâíåíèþ ñ øèðîêî èñïîëüçó-
ganization of the investigated tissue. The introduction åìûìè ìîíîñëîéíûìè êóëüòóðàìè. Íåäîñòàòêè è ïðåèìóùå
-ñòâà êàæäîãî òèïà êóëüòóð, à òàêæå èõ ïðèìåíåíèå â áèîëî-
of cells into the strange conditions leads to the
ãè÷åñêèõ è ìåäèöèíñêèõ èññëåäîâàíèÿõ ÿâëÿþòñÿ ïðåäìåòîì
impairment of intracellular interactions, polarization, ýòîãî îáçîðà.
distribution of receptors of growth factors and Êëþ÷åâûå ñëîâà: äâóõ- è òðåõìåðíîå êóëüòèâèðîâàíèå, ðå-
çèñòåíòíîñòü, äèôôåðåíöèðîâêà
hormones and makes its own input into the response of
cells to the tested factors. Recently the time of À. ². Õîðóæåíêî
cultivation in three-dimensional conditions is
considered to be one of the main pathways during the Äâî- ³ òðèâèì³ðíà êóëüòóðà êë³òèí
investigation of the properties of cells in vitro [52–54]. Ðåçþìå
Similar point of view is based on the statement that in Êóëüòèâóâàííÿ êë³òèí ññàâö³â çà òðèâèì³ðíèõ óìîâ ñòຠïð³-
vivo the cells in the organism are in the composition of îðèòåòíèì ó ÷èñëåííèõ á³îìåäè÷íèõ çàñòîñóâàííÿõ.  ãàëóç³
three-dimensional structures, for instance, thyroid òîêñèêîëî㳿 ³ ðîçðîáêè ïðîòèïóõëèííèõ ïðåïàðàò³â öå ïîâ’ÿ-
çàíî ç³ çíà÷íîþ ð³çíèöåþ ó â³äïîâ³ä³ êë³òèí, êóëüòèâîâàíèõ çà
follicles. It promotes the implementation of specific äâî- àáî òðèâèì³ðíèõ óìîâ, íà ïðîàïîïòè÷í³ ÷èííèêè. Êð³ì òî-
functions by tissues and organs. However, usually the ãî, âèÿâëåíî ³ñòîòí³ â³äì³ííîñò³ â ïîëÿðèçàö³¿ êë³òèí, ñòðóê-
investigation of these cultures raises a number of the òóð³ öèòîñêåëåòà, ðîçïîä³ë³ ðåöåïòîð³â äî øèðîêîãî ñïåêòðà
ãîðìîí³â, ðîñòîâèõ ÷èííèê³â ³ ò. ä. ó êë³òèíàõ ññàâö³â çàëåæíî
abovementioned methodological questions. On the â³ä óìîâ êóëüòèâóâàííÿ. Çðåøòîþ, öå ïðîÿâëÿºòüñÿ â ð³çí³é ðå-
other hand, in conditions of monolayer cultures the àêö³¿ êóëüòèâîâàíèõ êë³òèí íà ïîçàêë³òèíí³ ñòèìóëè. Íàéàäåê-
cells preserve many fundamental properties, inherent âàòí³øîþ ìîäåëëþ ðîñòó ñîë³äíèõ ïóõëèí in vitro íàðàç³ ââà-
æàþòü áàãàòîêë³òèíí³ ñôåðî¿äè. Êóëüòèâóâàííÿ ôîë³êóë³â
to them in vivo, including a part of antigens – ùèòîïîä³áíî¿ çàëîçè, àöèíóñ³â ìîëî÷íî¿ çàëîçè òà ³íøèõ
differentiation markers. Therefore, the most promising ñòðóêòóðíî-ôóíêö³îíàëüíèõ îäèíèöü, ùî çáåð³ãàþòü ïî÷àòêî-
method is deemed to be the use of the combination of âó îðãàí³çàö³þ òêàíèíè, äîçâîëÿº âèâ÷àòè ïîâåä³íêó, á³îõ³ì³÷í³
îñîáëèâîñò³ ³ ãåíåòè÷íèé ïðîô³ëü äèôåðåíö³éîâàíèõ êë³òèí. Ç
these two ways of cultivation during the investigation ³íøîãî áîêó, òðèâèì³ðí³ êóëüòóðè ìàþòü íèçêó îáìåæåíü ïî-
of adhesion, proliferation, migration, invasion, ð³âíÿíî ç øèðîêî âèêîðèñòîâóâàíèìè ìîíîøàðîâèìè êóëüòó-
differentiation of cells and many other processes, ðàìè. Íåäîë³êè é ïåðåâàãè êîæíîãî òèïó êóëüòóð, à òàêîæ ¿õ-
íº çàñòîñóâàííÿ â á³îëîã³÷íèõ ³ ìåäè÷íèõ äîñë³äæåííÿõ ñòà-
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