共查询到20条相似文献,搜索用时 15 毫秒
1.
Nikolay K. Popnikolov Jason Yang Raphael C. Guzman Steven M. Swanson Gudmundur Thordarson Gail Collins Frank Talamantes Satyabrata Nandi 《Journal of cellular physiology》1995,163(1):51-60
A new system for studying growth of normal human mammary epithelial cells in an in vivo environment using athymic nude mice is described. Human mammary epithelial cells dissociated from reduction mammoplasty specimens were embedded within collagen gels and subsequently transplanted subcutaneously into nude mice. Histological sections of recovered collagen gels showed epithelial cells arranged as short tubules with some branching. Proliferation of mammary epithelial cells was quantitated in vivo by 3 days' continuous infusion with 5 bromo-2′-deoxy-uridine followed by immunostaining of sections from recovered gels. Ovarian steroids administered to the host animals, resulting in blood serum levels normally found in the human female, had little or no effect on the proliferation of human mammary epithelial cells. Collagen gel embedded mouse mammary epithelial cells, mouse mammary explants, and host mammary glands all responded similarly to ovarian steroids, suggesting that the unresponsiveness of the human mammary epithelial cells under these conditions was not due to dissociation per se. However, an increased dose of 17β-estradiol or a growth factor combination containing epidermal growth factor, cholera toxin, and cortisol significantly stimulated the proliferation of human outgrowths. The growth factor response was dependent on the location of the cells, with the greatest response seen in the part of the gel proximal to the osmotic pump delivering the growth factors and the effect gradually waning in area more distal to the pump. The effect was especially striking since the mitotic figures could be easily identified and the labeling index was as high as 75%. The host mouse mammary gland also responded to growth factors, resulting in ductal hyperplasia. The proliferative and morphogenetic effects of various agents on normal human mammary epithelial cells embedded in collagen gel can be studied in vivo in nude mice. © 1995 Wiley-Liss, Inc. 相似文献
2.
Serum-free culture of enriched mouse anterior and ventral prostatic epithelial cells in collagen gel
Timothy Turner Howard A. Bern Peter Young Gerald R. Cunha 《In vitro cellular & developmental biology. Plant》1990,26(7):722-730
Summary Sustained growth of mouse ventral and anterior prostatic epithelial cells embedded within collagen gel matrix was achieved
in a serum-free medium composed of Dulbecco's modified Eagle's medium and Ham's F12 medium, 1∶1 (vol/vol), supplemented with
bovine serum albumin fraction V, epidermal growth factor, transferrin, cholera toxin, prolactin, 5α-dihydrotestosterone, cortisol,
putrescine, fibroblast growth factor, and a trace element mixture. Three-dimensional growth of prostatic epithelial cells
occurred inside the collagen gel matrix. This serum-free medium allowed cell growth greater than sevenfold over 10 d in culture.
Tissue recombination and cell culture techniques were integrated to demonstrate that cultured cells retained prostatic characteristics.
Following 10 d of culture, epithelial colonies from mouse ventral and anterior prostatic epithelial cell cultures were isolated
and combined with rat fetal urogenital sinus mesenchyme and grown for 4 wk under the renal capsule of intact athymic male
mice. These tissue recombinants showed distinctive prostatic histologic characteristic (alveoli and ducts lined with cuboidal
or columnar epithelium surrounded by stroma). When histologic sections of recombinants were stained with the Hoechst 33258,
epithelial cells of mouse origin were distinguishable from stromal cells of rat origin.
Aided by grants CA-05388 and CA-09041 from the National Institutes of Health, Bethesda, MD, and by M. A. R. C. fellowship
GM08730 to T. T. 相似文献
3.
Different mitogenic and phenotypic responses of human breast epithelial cells grown in two versus three dimensions 总被引:1,自引:0,他引:1
Jason Yang Arun Balakrishnan Susan Hamamoto Craig W. Beattie Tapas K. Gupta Sefton R. Wellings Satyabrata Nandi 《Experimental cell research》1986,167(2):563-569
Human breast epithelial cells, derived from fibroadenomas, were cultured under conditions promoting growth in two-dimensions (2D) as monolayers using the collagen-coated dishes and in three-dimensions (3D) inside the collagen gel matrix. Both epidermal growth factor (EGF) and cortisol (F) were required for maximal stimulation in 3D growth, but only cortisol was required for 2D growth. The growth stimulation of exogenously added type IV collagen was no greater than that of type I as a substrate in both the 2D and 3D growth. Immunocytochemical staining, using a polyclonal actin antibody, showed homogeneous staining in all cells in 2D monolayers, whereas more restricted distribution was observed in 3D outgrowths in the collagen gel matrix. The same cells, when cultured in 2D vs 3D, elicit different responses and the original phenotypes may be better maintained in 3D. 相似文献
4.
We have previously shown that isolated mouse fetal choroid plexus epithelial (CPE) cells penetrate a basement membrane matrix (Matrigel) substratein vitroto form single-layered epithelial vesicles embedded within the matrix. To determine which properties of the matrix are important for inducing or permitting cells to penetrate the substrate and organize into multicellular vesicles we have made quantitative changes to the basement membrane components and growth factors in cell cultures. Matrigel diluted to 33 or 10% with a collagen I gel was not permissive to cell invasion, and CPE cells formed a polarized epithelial monolayer on the substrate surface which had ultrastructural characteristics similar to those of CPE vesicles. Cells in these monolayers proliferated more rapidly than cells in epithelial vesicles. When deliberately embedded within a 33 or 10% Matrigel matrix, CPE cells were able to form vesicles, indicating that a dilute matrix is nonpermissive to cell invasion but promotes epithelial polarization and organization into vesicles. Cells embedded within a 100% collagen I matrix did not proliferate or form epithelial vesicles and the majority of cells did not remain viable. Addition of laminin to the collagen I gel promoted cell adhesion and cell survival, but did not promote the formation of extensive monolayers on the substrate nor the formation of epithelial vesicles within the matrix. Cell invasion into the 33% Matrigel matrix was induced by addition of laminin, nidogen, or a laminin–nidogen complex to the substrate or by addition of TGFβ2 to the culture medium, but not TGFβ1 or PDGF. These studies show that CPE cells are sensitive to quantitative changes in matrix composition, which influences their survival and proliferation and also their ability to penetrate the matrix and organize into multicellular epithelial vesicles. 相似文献
5.
Yasuhiro Tomooka Stephen E. Harris John A. McLachlan 《In vitro cellular & developmental biology. Plant》1985,21(4):237-244
Summary Epithelial cells from mouse seminal vesicles were enzymatically dissociated enriched by gradient centrifugation, and maintained
in collagen gel cultures with defined (serum-free) media. The epithelial origin of the cells was determined morhologically,
immunocytochemically, and biochemically. Cells formed three-dimensional colonies with a lumen in collagen gels. Cell number
was increased eight-fold within a 8 to 12-d culture period in a medium supplemented with epidermal growth factor (EGF) (10
ng/ml), insulin (10 μg/ml), transferrin (10 μg/ml), cholera toxin (10 ng/ml), and hydrocortisone (0.1 μg/ml). The cells required
eGF and insulin; the growth-promoting effects of these two peptide hormones were optimized by transferrin, cholera toxin,
and hydrocortisone. Fetal bovine serum did not support growth; rather, it suppressed the stimulated growth observed in serum-free
media. A time-course study revealed that a lag period preceded rapi growth. The collagen gel, serum-free culture provides
a powerful tool to study the effects of hormones on proliferation and differentiation of androgen sensitive cells. 相似文献
6.
《Cell biology international reports》1982,6(10):969-975
Collagen gel matrix has been used successfully to promote sustained growth of human normal mammary epithelial cells in primary culture using serum-containing medium supplemented with hormones and growth factors (Nandi et al., 1932). Sustained growth can now be accomplished in a serum-free medium consisting of a 1:1 mixture of Ham's F12 and DMEM supplemented with insulin, transferrin, epidermal growth factor, cholera toxin, cortisol, and BSA. Human normal mammary epithelial cells derived from reduction mammoplasties can be routinely propagated in this serum-free medium. The extent of growth and the resulting three-dimensional outgrowths in this serum-free medium, using the collagen gel matrix system, are comparable to those seen in serum-containing medium. This is the first demonstration of sustained growth of human normal mammary epithelial cells in serum-free primary culture. 相似文献
7.
Epithelial cells were isolated from the ventral prostate gland of the mouse after prolonged incubation in a mixture of collagenase, Dispase and hyaluronidase followed by extensive pipetting. The isolated epithelial cells were then embedded in collagen gels. After cultivation in Dulbecco's modified Eagle's medium supplemented with fetal bovine serum, epidermal growth factor, 5 alpha-dihydrotestosterone and cortisol, stimulation of growth and branching morphogenesis of the epithelial cells were observed. Under these culture conditions, growth of contaminating fibroblastic cells was rarely seen. These observations suggest that hormones including androgen directly stimulate the growth and morphogenesis of mouse prostate epithelial cells in culture. 相似文献
8.
Growth of mouse vaginal epithelial cells in culture: Effect of sera and supplemented serum-free media 总被引:3,自引:0,他引:3
Taisen Iguchi Francis-Dean A. Uchima Howard A. Bern 《In vitro cellular & developmental biology. Plant》1987,23(8):535-540
Summary Normal mouse vaginal epithelial cells isolated from ovariectomized ca. 35-d-old BALB/cCrgl mice were grown in primary culture
using collagen gel metrix and a serum-free medium composed of a 1∶1 mixture of Dulbecco’s modified Eagle’s medium and Ham’s
F12 (D:H) medium supplemented with insulin (IN), epidermal growth factor (EGF), cholera toxin, transferrin, and bovine serum
albumin V (BSA). Three-dimensional cellular outgrowths occurred inside the collagen gel matrix. The contribution of each factor
to cell growth was examined by individual addition to the basic D:H medium and by individual deletion from the complete serum-free
medium. When added individually, only IN promoted growth. Deletion of IN from the complete serum-free medium markedly, diminished
growth; deletion of EGF or BSA slightly diminished growth. When horse, fetal bovine, or chicken serum was added to the basal
D:H medium, only with increasing doses of horse serum was there enhanced cell growth. The effect of 17?-estradiol and diethylstilbestrol
on the growth of cells was also tested, using a suboptimal medium of D:H supplemented with BSA and IN, or a minimal medium
supplemented with IN alone. During the 8-d time period, addition of estrogen did not enhance cell growth in either medium.
To date, we have been unable to demonstrate a mitogenic effect of estrogen; rather a dose-dependent inhibition of proliferation
is seen.
This investigation was supported by grants CA-05388 and CA-09041, awarded by the National Institutes of Health, DHHS, Bethesda,
MD. 相似文献
9.
E. Jane Ormerod Philips S. Rudland 《In vitro cellular & developmental biology. Plant》1988,24(1):17-27
Summary The mechanism of induction of tubular outgrowths in vitro on floating collagen gels and the influence of extracellular factors
on this process have been investigated using the clonal rat mammary epithelial cell line, Rama 25. Growth of Rama 25 on such
floating gels causes their contraction. Contraction of the gel is accompanied by a 10-fold increase in the number of cells
per unit area, a change in cell shape, and a convolution of the epithelial cell sheet. Gels folded over manually show an 11-times
higher incidence of tubules along the folds than on the flat surface. Tubular formation begins when cords of cells develop
from local proliferations of the cell sheet and become canalized. Tubules follow wrinkles in the gel and branch to yield monopodial,
dichotomous, or lobular architecture.
Hydrocortisone and insulin, in the presence of serum, stimulate both narrow and thick tubular structures on folded gels, whereas
extra additions of 1 ng/ml cholera toxin or 100 ng/ml epidermal growth factor preferentially stimulate thick tubular structures.
Floating glutaraldehyde-fixed gels, very thick collagen gels, and collagen gels prepared on the top of rigid steel grids fail
to support the formation of tubules, suggesting that flexibility and access of the medium to basal surfaces are important
to their genesis. Incorporation of hyaluronic acid into the gel matrix preferentially inhibits the thick tubular outgrowths.
Thus, the branching tubular structures generated by Rama 25 can be influenced in different ways by various extracellular factors
in the medium and in the gel.
During the course of this work E. J. Ormerod was in receipt of a Ludwig Research Studentship. 相似文献
10.
Elham Hasanzadeh Somayeh Ebrahimi-Barough Narges Mahmoodi Amir Mellati Houra Nekounam Arefeh Basiri Shiva Asadpour Diba Ghasemi Jafar Ai 《Cell biology international》2021,45(1):140-153
Human endometrial stem cells (hEnSCs) that can be differentiated into various neural cell types have been regarded as a suitable cell population for neural tissue engineering and regenerative medicine. Considering different interactions between hormones, growth factors, and other factors in the neural system, several differentiation protocols have been proposed to direct hEnSCs towards specific neural cells. The 17β-estradiol plays important roles in the processes of development, maturation, and function of nervous system. In the present research, the impact of 17β-estradiol (estrogen, E2) on the neural differentiation of hEnSCs was examined for the first time, based on the expression levels of neural genes and proteins. In this regard, hEnSCs were differentiated into neuron-like cells after exposure to retinoic acid (RA), epidermal growth factor (EGF), and also fibroblast growth factor-2 (FGF2) in the absence or presence of 17β-estradiol. The majority of cells showed a multipolar morphology. In all groups, the expression levels of nestin, Tuj-1 and NF-H (neurofilament heavy polypeptide) (as neural-specific markers) increased during 14 days. According to the outcomes of immunofluorescence (IF) and real-time PCR analyses, the neuron-specific markers were more expressed in the estrogen-treated groups, in comparison with the estrogen-free ones. These findings suggest that 17β-estradiol along with other growth factors can stimulate and upregulate the expression of neural markers during the neuronal differentiation of hEnSCs. Moreover, our findings confirm that hEnSCs can be an appropriate cell source for cell therapy of neurodegenerative diseases and neural tissue engineering. 相似文献
11.
Several pathological and disease conditions can alter the mechanical properties of the extracellular matrix (ECM). Conversely,
some diseases may arise from changes in the density or rigidity of the ECM. This necessitates the use and development of in vitro models to understand how both biophysical and biochemical signals regulate complex cellular behaviors. T47D breast epithelial
cells will differentiate into duct-like tubules when cultured in a floating three-dimensional (3D) collagen gel, but not a
3D collagen gel that is left attached to the culture dish. This paper details several protocols we have developed for analyzing
breast cell biology in 3D matrices, including culturing cells in 3D collagen gels, immunostaining cellular structures, and
performing biochemical procedures directly from cells embedded in collagen gels. 相似文献
12.
Mouse submandibular salivary epithelial cell growth and differentiation in long-term culture: Influence of the extracellular matrix 总被引:7,自引:0,他引:7
Elisa M. Durban 《In vitro cellular & developmental biology. Plant》1990,26(1):33-43
Summary The adult mouse submandibular salivary gland provides a good model system to study gene regulation during normal and abnormal
cell behavior because it synthesizes functionally distinct products ranging from growth factors and digestive enzymes to factors
of relevance to homeostatic mechanisms. The present study describes the long-term growth and differentiation of submandibular
salivary epithelial cells from adult male mice as a function of the culture substratum. Using a two-step partial dissociation
procedure, it was possible to enrich for ductal cells of the granular convoluted tubules, the site of epidermal growth factor
synthesis. Long-term cell growth over a period of 2 to 3 mo. with at least 3 serial passages was obtained only within three-dimensional
collagen gels. Cells grew as ductal-type structures, many of which generated lumens with time in culture. Electron microscopic
analysis in reference to the submandibular gland in vivo revealed enrichment for and maintenance of morphologic features of
granular convoluted tubule cells. Reactivity with a keratin-specific monoclonal antibody established the epithelial nature
of the cells that grew within collagen. Maintenance of cell differentiation, using immunoreactivity for epidermal growth factor
as criterion, was determined by both cytochemical and biochemical approaches and was found to be dependent on the collagen
matrix and hormones. Greater than 50% of the cells in primary collagen cultures contained epidermal growth factor only in
the presence of testosterone and triiodothyronine. In contrast, cells initially seeded on plastic or cycled to plastic from
collagen gels were virtually negative for epidermal growth factor. Biochemical analysis confirmed the presence of a protein
with an apparent molecular weight of 6000 which comigrated with purified mouse epidermal growth factor. Epidermal growth factor
was also present in detectable levels in Passage 1 cells. This culture system should permit assessment of whether modulation
of submandibular gland ductal cell growth can be exerted via a mechanism that in itself includes epidermal growth factor and
its receptor and signal transduction pathway.
This work was supported by Public Health Service grant DE07766 from the National Institute of Dental Research, National Institutes
of Health, Bethesda, MD. 相似文献
13.
17β-Estradiol has been demonstrated to protect blood-brain barrier from disruption and attenuate brain injury in various conditions. The aim of this study was to investigate the effect of 17β-estradiol on the blood-retina barrier (BRB) breakdown induced by intravitreous injection of vascular endothelial growth factor (VEGF), a significant mediator of vascular permeability. Intravitreous injection of VEGF was performed to initiate BRB breakdown in male rats with PBS in the contralateral eye as control. 2 doses of 17β-estradiol and vehicle control were given to 3 groups of rats. The integrity of the BRB was quantified by Evans blue technique and assessed by fluorescent dyes in retinal sections and wholemounts. BRB breakdown was achieved by VEGF as retinal vascular permeability was increased compared with control eyes (14.66±4.09 vs. 4.94±1.20 μl/g/h, p<0.01). Vascular permeability in the 2 groups treated with 17β-estradiol was reduced compared with control (14.66±4.09 vs. 10.26±3.67 vs. 7.37±2.22 μl/g/h, p<0.01). Rhodamine isothiocyanate (RhIC) extravasation in retinal sections and Evans blue-albumin complex leakage in retinal wholemounts were also decreased in the 2 treatment groups. These results suggest that 17β-estradiol attenuates BRB breakdown induced by VEGF in male rats, which may provide a new role of 17β-estradiol in ocular diseases. 相似文献
14.
Growth requirements and characterization of rat cervical epithelial cells in culture 总被引:1,自引:0,他引:1
The extended culture of rat cervical epithelial cells can be achieved in the absence of a fibroblast feeder layer by utilizing collagen gels and a complex growth medium. The medium contains a 1:1 mixture of RPMI-1640 and Ham's F12 supplemented with 7.5% porcine serum and epidermal growth factor, cholera toxin, transferrin, insulin, and hydrocortisone. Under these culture conditions the cells show rapid log-phase growth and high saturation densities while retaining the ultrastructural characteristics of immature squamous metaplastic cells of the rat uterine cervix even after extended passage. In a manner similar to epithelial cells from a variety of sources, rat cervical epithelial cells form hemicysts at confluence in vitro when cultured on impermeable substrates. The development of these methods for culturing cervical epithelial cells provides an experimental system for the study of factors important in regulating the growth and differentiation of metaplastic squamous epithelial cells. 相似文献
15.
Human breast epithelial cells in serum-free collagen gel primary culture: growth, morphological, and immunocytochemical analysis 总被引:2,自引:0,他引:2
J Yang A Balakrishnan S Hamamoto J J Elias W Rosenau C W Beattie T K Das Gupta S R Wellings S Nandi 《Journal of cellular physiology》1987,133(2):228-34, 254-5
Human breast epithelial cells derived from various sources (fibroadenoma, reduction mammoplasty, and mastectomy tissues from premenopausal patients) have been cultured in collagen gel matrix using serum-free medium. Response to various additives has been analyzed for growth-promoting effect when added to a basal medium containing insulin, cholera toxin, and BSA. A consistent observation has been the effect of EGF and cortisol in growth stimulation of human breast epithelial cells, while separately, each additive elicited only a small response. Under this condition, employing EGF and cortisol combinations, these cells gave rise to organized colonies consisting of clusters of cells, usually spherical, without any duct-like extensions. Ultrastructural and immunocytochemical studies, using a panel of monoclonal and polyclonal antibodies, have shown that cell types and features that can be identified in the original breast tissue can also be delineated in the progeny populations. The topographical feature, consisting of lumina surrounded by a single inner layer of epithelial cells and an outer layer of basal/myoepithelial cells, can be re-created in the collagen gel system starting from small clumps of cells. 相似文献
16.
Autophagy is a catabolic process providing an alternative energy source for cells under stressful conditions such as starvation, growth factor deprivation or hypoxia. During involution of the bovine mammary gland autophagy is induced in mammary epithelial cells (MECs) as a survival mechanism, and is tightly regulated by hormones and growth factors necessary for gland development. In the present study we adapted the three-dimensional culture model to investigate the role of autophagy during formation of alveoli-like structures by bovine BME-UV1 MECs grown on extracellular matrix (ECM) components. Using confocal microscopy and Western-blot analyses of autophagic and apoptotic markers: LC3, and cleaved caspase-3, we showed that autophagy was induced in centrally localized cells within the developing acini. These cells lacked a direct contact with ECM, and formed a distinct population from the outer layer of cells. Induction of autophagy preceded apoptosis, but did not inhibit the formation of a hollow lumen. In the presence of steroid hormones: 17β-estradiol and progesterone, although autophagy was augmented, acini formation proceeded normally. In contrast, the major lactogenic hormone: prolactin, which supports functional differentiation of alveoli, did not alter induction of autophagy within the spheroids. BME-UV1 cells cultured on Matrigel in the presence of growth factors IGF-I and EGF formed larger, underdeveloped acini without lumens due to caspase-3 inhibition, and sustained autophagy in the centre of the spheroids, while TGF-β1 accelerated apoptosis, and increased autophagy significantly. Our observations suggest that sex steroids 17β-estradiol and progesterone, as well as growth factor TGF-β1 may regulate the development of the bovine mammary gland by inducing autophagy in addition to regulating proliferation and apoptosis of MECs. These data indicate that autophagy may play an important role during alveolargenesis. 相似文献
17.
Ziyad Jabaji Garrett J. Brinkley Hassan A. Khalil Connie M. Sears Nan Ye Lei Michael Lewis Matthias Stelzner Martín G. Martín James C. Y. Dunn 《PloS one》2014,9(9)
Background
We previously reported in vitro maintenance and proliferation of human small intestinal epithelium using Matrigel, a proprietary basement membrane product. There are concerns over the applicability of Matrigel-based methods for future human therapies. We investigated type I collagen as an alternative for the culture of human intestinal epithelial cells.Methods
Human small intestine was procured from fresh surgical pathology specimens. Small intestinal crypts were isolated using EDTA chelation. Intestinal subepithelial myofibroblasts were isolated from a pediatric sample and expanded in vitro. After suspension in Matrigel or type I collagen gel, crypts were co-cultured above a confluent layer of myofibroblasts. Crypts were also grown in monoculture with exposure to myofibroblast conditioned media; these were subsequently sub-cultured in vitro and expanded with a 1∶2 split ratio. Cultures were assessed with light microscopy, RT-PCR, histology, and immunohistochemistry.Results
Collagen supported viable human epithelium in vitro for at least one month in primary culture. Sub-cultured epithelium expanded through 12 passages over 60 days. Histologic sections revealed polarized columnar cells, with apical brush borders and basolaterally located nuclei. Collagen-based cultures gave rise to monolayer epithelial sheets at the gel-liquid interface, which were not observed with Matrigel. Immunohistochemical staining identified markers of differentiated intestinal epithelium and myofibroblasts. RT-PCR demonstrated expression of α-smooth muscle actin and vimentin in myofibroblasts and E-Cadherin, CDX2, villin 1, intestinal alkaline phosphatase, chromogranin A, lysozyme, and Lgr5 in epithelial cells. These markers were maintained through several passages.Conclusion
Type I collagen gel supports long-term in vitro maintenance and expansion of fully elaborated human intestinal epithelium. Collagen-based methods yield familiar enteroid structures as well as a new pattern of sheet-like growth, and they eliminate the need for Matrigel for in vitro human intestinal epithelial growth. Future research is required to further develop this cell culture system for tissue engineering applications. 相似文献18.
Effect of hormones and EGF on proliferation of rat mammary epithelium enriched for alveoli : An in vitro study 总被引:4,自引:0,他引:4
Virgin rat mammary epithelium enriched for alveoli were embedded in a collagen gel matrix to study the direct effect of mammogenic hormones and epidermal growth factor (EGF) on their growth over a 12-day culture period. Serum-supplemented medium alone caused a 3- to 4-fold increase in cell number, whereas medium containing insulin, prolactin, progesterone, cholera toxin and serum caused a 15-fold increase. Cultures resulting from this substantial cell number increase consisted of large, smooth-bordered epithelial colonies with relatively few (< 1%) single cells surrounding them. An equal increase in cell number was obtained when progesterone was replaced by hydrocortisone in the above-mentioned medium, but these cultures contained predominantly single spindle-shaped cells with a few small epithelial colonies. The smooth-bordered epithelial colonies consisted solely of mammary epithelial cells, since they contained thioesterase II, an enzyme found exclusively in mammary epithelium. The identity of the single spindle-shaped cells remains to be determined. The addition of EGF to serum or serum, hormone and cholera toxin-supplemented medium did not enhance the proliferative effect of these factors on the alveolar-enriched population. 相似文献
19.
Substitution for mesenchyme by basement-membrane-like substratum and epidermal growth factor in inducing branching morphogenesis of mouse salivary epithelium 总被引:11,自引:0,他引:11
Mouse salivary epithelium cannot undergo branching morphogenesis in the absence of the surrounding mesenchyme. To clarify the nature of the mesenchymal influence on the epithelium, we have investigated the culture conditions in which the epithelium could normally branch in the absence of mesenchymal cells. Combination of basement-membrane-like substratum (Matrigel) and epidermal growth factor (EGF) could substitute for the mesenchyme, the epithelium showing typical branching morphogenesis. Transforming growth factor alpha had the same effect as EGF. Matrigel plus basic fibroblast growth factor or transforming growth factor beta 1 and collagen gel plus EGF were not sufficient to support the branching of the epithelium. These results clearly reveal that the role of mesenchyme in salivary morphogenesis is both to provide the epithelium with an appropriate substratum and to accelerate growth of the epithelium. 相似文献
20.
Debabrata Ghosh Keith G. Danielson James T. Alston Susan Heyner 《In vitro cellular & developmental biology. Animal》1991,27(9):713-719
Summary Epithelial cells were isolated from mouse endometrium and cultured on two types of extracellular matrix, namely, rat-tail
collagen (type I) gels and basement membrane extract (BME) derived from the Engelbreth-Holm-Swarm murine sarcoma. Cell attachment
in serum-free medium during the initial 24 h after seeding was approximately twofold higher on BME compared with collagen
type I. Addition of serum to the medium enhanced cell attachment on both matrices. On both collagen and BME, uterine cells
grew as smooth-bordered colonies, and within a week of culture the cells became cuboidal to columnar in shape. Electron microscopy
revealed the presence of apical microvilli associated with a glycocalyx, junctional complexes, tonofilaments, short strands
of undilated endoplasmic reticulum, Golgi complex, and lipid droplets. However, cells on BME showed a higher degree of differentiation
as assessed by occasional formation of small patches of basement membranelike structure subjacent to the flattened basal surface
and formation of glandlike structures within the matrix. Proliferation of these cells as measured by radioactive thymidine
incorporation into DNA was increased threefold by addition of epidermal growth factor (EGF) and insulin to the medium, but
was not changed by 17β-estradiol. The expression of progesterone receptors by uterine epithelial cells grown on both matrices was doubled by addition
of EGF and estradiol to the medium.
This work was supported in part by a Rockefeller Foundation postdoctoral fellowship (D.G.), and NIh grant 23511. 相似文献