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1.
Neil C. Talbot Thomas J. Caperna 《In vitro cellular & developmental biology. Animal》1998,34(10):785-798
Summary Secondary culture of nontransformed bile duct epithelium has been difficult to achieve. STO feeder cell-dependent secondary
cultures of adult pig bile duct cells were established from primary cultures of adult pig liver cells. Adult pig hepatocytes
exhibited limited or no replication and were lost from the secondary culture at Passage 3 or 4. In contrast, adult pig bile
duct cells replicated and were carried for 4–8 passages in secondary culture. A simple method to produce nearly pure pig intrahepatic
bile duct cultures was first to freeze a relatively crude liver cell preparation. Upon subsequent thawing, all hepatocytes
and most macrophages were lysed. Bile duct cells composed 95% of the surviving cells after the freeze/thaw, and they grew
out rapidly. The bile duct cells grew on top of the STO feeder cells as closely knit epithelial, colonial outgrowths. Histocytochemical
and biochemical analyses demonstrated high levels of gamma-glutamyltranspeptidase activity and low levels of P450 activity
in the bile duct cultures. The bile duct cells spontaneously adopted a multicellular ductal morphology after 7–10 d in static
culture which was similar to that found in in vivo pig liver. Transmission electron microscopic examination revealed complex
junctions and desmosomes typical of epithelium, and lumenally projecting cilia typical of in vivo intrahepatic bile ductules.
This simple method for the coculture of pig intrahepatic bile duct cells which adopt in vivo-like structure may facilitate
biological studies of this important, but difficult to culture, cell type. 相似文献
2.
Effects of medium and substratum conditions on the rates of DNA synthesis in primary cultures of bile ductular epithelial cells 总被引:4,自引:0,他引:4
Georg A. Mathis Alphonse E. Sirica 《In vitro cellular & developmental biology. Plant》1990,26(2):113-118
Summary Select medium and substratum conditions were investigated for their effects on semiconservative DNA synthesis in essentially
pure primary cultures of bile ductular epithelial cells that were initially isolated from cholestatic rat livers at 6 to 10
wk after bile duct ligation. DNA synthesis in these cultured cells was serum-dependent, being at its highest level when the
concentration of fetal bovine serum present in the medium was maintained at 10%. This serum-dependent DNA synthesis was completely
inhibited when 10 mM hydroxyurea was also included in the medium, and bile ductular cells cultured in the continued presence of 1.0% fetal bovine
serum showed only marginal DNA synthesis during 8 to 10 d of primary culture when compared with no-serum controls. Maximum
rates of serum-dependent DNA synthesis were obtained when the bile ductular cells were cultured for 7 to 14 d on tissue culture
plastic coated with obtained when the bile ductular cells were cultured for 7 to 14 d on tissue culture plastic coated with
either fibronectin from bovine plasma or type I rat-tail collagen. Cells cultured on plastic coated with basement membrane
Matrigel exhibited the lowest levels of DNA synthesis, whereas those on plastic alone had intermediate amounts. Furthermore,
the addition of epidermal growth factor (50 ng·ml−1·d−1) to medium supplemented with 1.0% fetal bovine serum greatly enhanced the rate of DNA synthesis in bile ductular cells after
6 d in primary culture on type I collagen-coated plastic over that measured in solvent control cultures. These findings indicate
that our bile ductular epithelial cell culture model is potentially useful in the study of biliary cell growth regulation
and carcinogenesis.
This investigation was supported by USPHS grant RO1 CA 39225 to A. E. Sirica by the National Cancer Institute, Department
of Health and Human Services, Bethesda, MD. During the period of this study, G. A. Mathis was a recipient of a Fellowship
from the Fund for Academic Career Development of the State of Zurich, Switzerland. 相似文献
3.
Lopez-Souza N Avila PC Widdicombe JH 《In vitro cellular & developmental biology. Animal》2003,39(7):266-269
Airway epithelial cultures are generally derived from tracheas postmortem or from surgical specimens of nasal polyps or turbinates. Scrapings of the mucosal surface have been little used as starting material for cultures because of their low yield of epithelial cells and their contamination with mucous secretions, blood, and underlying connective tissue. For the first time, we report that human airway epithelial cells obtained from nasal scrapings or bronchial brushings can be grown in culture to produce polarized cell sheets suitable for studies of vectorial transport. 相似文献
4.
E. M. Earley S. C. Finley W. H. Finley L. Y. F. Hsu H. J. Kim J. C. Petricciani P. C. Vinson 《In vitro cellular & developmental biology. Plant》1976,12(9):639-642
Summary Cell cultures were established from the biopsies of lung, skin and kidney from each of nine human fetuses, and chromosome
analyses were performed on material through the fifth subculture. Kidney cell cultures generally showed a higher level of
polyploidy than lung or skin. The frequencies of hyperdiploid cells and those with structural abnormalities were consistent
with the low levels found in cultures of human lymphocytes. The data provide a normal cytogenetic baseline for human fetal
material which may be useful in a variety of studies.
Supported by Food and Drug Administration contracts FDA 74-51 and 74-52.
Authors are listed alphabetically. 相似文献
5.
James H. Resau Ph.D. John R. Cottrell Kathryn A. Elligett Eric A. Hudson 《Cell biology and toxicology》1987,3(4):441-458
Human esophageal, tracheal, and pancreatic ductal fragments were collected at autopsy after a postmortem interval of 12 hours or less and maintained in explant organ culture for 30 days. The viability and growth of the explants was assessed by morphology, LDH enzyme release, and cellular outgrowth. The viability and growth of the bronchial explant epithelium was directly related to the postmortem interval. Esophageal epithelial regeneration followed the desquamation of the superficial cell layers. Pancreatic epithelia appeared to grow more slowly and with less outgrowth than the other tissues. Epithelial cell growth along the explant surface and onto the culture dish appeared to proceed through the well-characterized process that follows cell injury, i.e., flattening, migration, replication, and differentiation. Thus, sufficient numbers of viable epithelial cells capable of regeneration were present in routine autopsy epithelium, but there was considerable variation from tissue to tissue and case to case. The most effective and accurate approach to follow when evaluating and predicting the growth and viability of these explants is by using a combination of morphologic, enzymatic and biologic assays. Errors in the interpretation of viability are possible when only one assay method is utilized. These tissues grown in explant organ culture are suitable for studies on the mechanism and response of epithelia to cell injury, recovery and wound healing.Abbreviations 4F-1G
4% formaldehyde, 1% glutaraldehyde
- HIFBS
heat inactivated fetal bovine serum
- IA
immediate autopsy
- LDH
lactate dehydrogenase
- OsO4
osmium tetroxide
- RA
routine autopsy 相似文献
6.
R. Joplin A. J. Strain J. M. Neuberger 《In vitro cellular & developmental biology. Plant》1989,25(12):1189-1192
Summary Biliary epithelial cells (BEC) lining the intra-hepatic biliary ducts are the site of damage in several immunologically mediated
liver diseases. BEC are difficult to isolate since they represent only 5% of the total cell number in normal liver. In this
communication, a novel method for their isolation from normal liver is presented using a monoclonal antibody (HEA125) with
specificity for an epithelial cell surface glyco-protein reported to be expressed in liver only by biliary epithelium. By
combining differential density centrifugation and immuno-magnetic separation using HEA125 pure BEC (105 cells/g fresh tissue) were prepared routinely. These cells were maintained in culture for up to 4 weeks with significant
increases in cell numbers. The ability to prepare BEC from human liver offers an opportunity to develop In Vitro models to
investigate the aetiology of diseases in intra-hepatic biliary epithelium.
EDITOR’S STATEMENT This is a novel application to purification of specific liver cell types directly from tissue. It is well-suited
for rapid communication because of its novelty and potential utility to investigators. 相似文献
7.
Primary liver cell cultures grown on gas permeable membrane as source for the collection of primary bile 总被引:2,自引:0,他引:2
Ernst Petzinger Wolfram Föllmann Helmut Acker Joachim Hentschel Karl Zierold Rolf K. H. Kinne 《In vitro cellular & developmental biology. Plant》1988,24(6):491-499
Summary Isolated rat hepatocytes maintained in primary culture on gas permeable membrane for 20 h form monolayers and establish at
their cell borders a network of canaliculi (approximate diameter 3.5 μm). In the presence of the known choleretic bile acid
dehydrocholate, dilation of canaliculi occurs. When nonfluorescent carboxyfluorescein diacetate ester is added to the culture
medium, fluorescent carboxyfluorescein appears in the intracanalicular space. In the dilated state, fluid containing the fluorescent
compound could be collected from the canaliculi by puncture with a micropipette. The intracanalicular space shows a negative
electrical potential difference of 31 mV in reference to the bath solution and is 13.5 mV more positive with reference to
recordings from the cytosol of cultured rat hepatocytes. Cultured rat hepatocytes grown on gas permeable membrane are energetically
stable over 3 d. On Day 4, ATP levels increase markedly, whereas Na+−K+-ATPase activity declines. Ionic composition of hepatocytes, as measured by electronprobe element analysis on cryosection
samples, does not change markedly during monolayer formation. With formation of bile canaliculi, the activity of alkaline
phosphatase rapidly increases within 24 h and is stable for the next 3 d. Within that time the activity of γ-glutamyltranspeptidase,
however, increases steadily, reaching a 1.6-fold higher activity than freshly isolated hepatocytes. Bile acids appear in the
culture supernatant after 1 d. When unconjugated [14C]cholic acid is added to the cultures the supernatant contains also [14C]tauro- and [14C]glycocholic acid, indicating the preservation of conjugation capacity in these cultures. Total bile acid concentrations
in the supernatant increase from 5 to 26 μM on Day 4. The cultures do not secrete α-fetoprotein. Monolayer cultures of hepatocytes in the presence of choleretic bile
acids seem to be a suitable model system to collect and to analyze the composition of primary bile. In conjunction with the
electrical parameters, it is possible to describe directly properties of bile secretion at the canalicular pole of the intact
hepatocyte.
This work was supported by the Deutsche Forschungsgemeinschaft, grant no. PE 250/5-1. 相似文献
8.
Gary D. Stoner Curtis C. Harris David G. Bostwick Raymond T. Jones Benjamin F. Trump Elizabeth W. Kingsbury Elliott Fineman Carnell Newkirk 《In vitro cellular & developmental biology. Plant》1978,14(7):581-590
Summary Epithelial cells derived from bovine pancreatic duct have been grown continuously in culture for 30 weeks (approximately 90
doublings of the cell population). The cells were grown in Eagle's minimal essential medium supplemented with 10% heat-inactivated
fetal bovine serum, 2 mM glutamine, 0.1 mM nonessential amino acids, and antibiotics. In confluent cultures, the cells are
multilayered and form circular structures. When tested at various passages, the cells neither formed colonies in soft agar
nor produced tumors after inoculation into athymic, nude mice. Hydrocortisone (1 and 5 μg per ml) and insulin (1,5 and 10
μg per ml) had no effect on the growth of the cells. β-Retinyl acetate inhibited growth rate and cell yield at a concentration
of 5 μg per ml but was not growth-inhibitory at lower concentrations. By electron microscopy the cells have numerous mitochondria,
Golgi and microvilli. Mucous droplets were observed in a small proportion of the cells. Desmosome-like structures and occluding
junctions were observed more frequently between cells that had been transferred as aggregates than between cells transferred
as single cells. Cytochemical studies indicated that some cells produce PAS positive granules that were not removed after
treatment of the cultures with diastase. Eleven cell clones were isolated from the mass culture. The growth rates of the clones
are different as well as the period of time in which the clones can be propagated in vitro.
This work was supported in part by Y01 CP 60204 and N01 CP 43237. 相似文献
9.
Sherwood Githens Jane A. Schexnayder Kemlesh Desai Christina L. Patke 《In vitro cellular & developmental biology. Plant》1989,25(8):679-688
Summary Interlobular duct fragments from the pancreas of the rat were isolated by collagenase digestion and filtration, embedded in
a matrix of rat-tail collagen, and cultured in a 1∶1 mixture of Dulbecco’s minimal essential and Ham’s F12 media supplemented
with cholera toxin (CT, 100 ng/ml) and epidermal growth factor (EGF, 10 ng/ml) in addition to supplements used previously,
thereby improving the yield of ducts by a factor of two compared with previous resuts. The ducts were harvested by digestion
of the collagen matrix with collagenase and were then dissociated by treatment with EDTA in divalent cation-free salt solution,
followed by digestion with collagenase and hyaluronidase. The resulting tissue fragments were suspended in collagen and cultured
as were the ducts. Numerous cysts appeared as a function of time and some of these enlarged dramatically. Some of the larger
cysts exhibited secondary tubular processes extending into the surrounding collagen. The addition of bovine pituitary extract
(BPE, 50 μg/ml) doubled the number of cysts, whereas omission of serum or CT+EGF reduced the number. BPE or forskolin could
substitute effectively for CT. Agents that stimulate (secretin) or inhibit (e.g., ouabain or acetazolamide) fluid-electrolyte
secretion in vivo had no effect on the number or average diameter of the cysts. The cysts were 83 to 88% epithelial with the
balance of the cells being fibroblastic in appearance. Some cysts consisted only of epithelium. The proliferative capacity
of the cystic epithelium was shown, by the presence of mitotic figures and by an autoradiographic labeling index of 22 to
30% after a 24-h exposure to [3H]thymidine. The labeling index was reduced by the omission of CT+EGF. Transmission electron microscopy showed that the cysts
exhibited morphologic features of duct epithelium in vivo, including apical microvilli, lateral, interdigitations of the plasma
membrane, and typical cytoplasmic organelles. 相似文献
10.
Kazuhiro Takeuchi Ikuro Maruyama Sinichi Yamamoto Toshimichi Oki Yukihiro Nagata 《In vitro cellular & developmental biology. Animal》1991,27(9):720-724
Summary In the present study we have established a pure monolayer culture system of human fallopian tube epithelial cells. The cells
were isolated using collagenase digestion, and were cultured in Medium 199 supplemented with 15% fetal bovine serum. The epithelial
cells derived from primary and secondary culture were characterized using immunocytochemical staining and electron microscopy.
The cells continued to grow for 2 to 3 wk once the monolayer culture of the cells was established. It is currently possible
to maintain the cultures until the third generation. Proliferation of these cells was enhanced by epidermal growth factor
but not by basic-fibroblast growth factor, insulin, transferrin, estradiol-17β, or progesterone. This culture system offers a good model for determining characteristics of the tubal epithelium and would
permit effective study of co-culture with embryos. 相似文献
11.
Peter K. A. Jensen Aage J. Therkelsen 《In vitro cellular & developmental biology. Plant》1982,18(10):867-871
Summary Overgrowth with fibroblasts has been a major problem in the cultivation of normal human skin epithelium. In the present study
it is shown that the addition of spermine to the culture medium in micromolar concentrations has a differential cytotoxic
effect on fibroblasts allowing the cultivation of human skin epithelial cells in primary culture without fibroblastic overgrowth.
Putrescine, another polyamine, is shown to be equally cytotoxic to fibroblasts and epithelial cells when added in millimolar
concentrations; below this concentration range no cytotoxic effect could be demonstrated. This difference in cytotoxicity
between spermine and putrescine is suggested to depend on the conversion of spermine, but not putrescine, and to highly cytotoxic
products by an amine oxidase present in fetal bovine serum.
This project was supported by the Novo foundation. 相似文献
12.
Vernon E. Steele Julia T. Arnold 《In vitro cellular & developmental biology. Plant》1985,21(12):681-687
Summary Nasal turbinate epithelial cells were isolated from rats, rabbits, and humans using either a surgical or an in situ enzyme
incubation technique. The culture conditions that permit optimal cell attachment and selective growth of the nasal epithelial
cells were determined. These conditions will permit the long-term culture of these cells where typically 20 to 30 population
doublings were observed. Differences between rat and human nasal epithelial cells were seen in substrate requirements, colony-forming
efficiency, and response to fetal bovine serum and bovine serum albumin. These methodology and results will permit mechanistic
studies of normal and abnormal cellular function and comparative response studies between nasal epithelial cells from rats
and humans.
This work was supported under U.S. Environmental Protection Agency contract 68-02-4032. 相似文献
13.
N. Auersperg C. H. Siemens S. E. Myrdal 《In vitro cellular & developmental biology. Plant》1984,20(10):743-755
Summary The ovarian surface epithelium (OSE) represents a minute fraction of the cell mass of the ovary but gives rise to over 80%
of human ovarian carcinomas. No experimental models for the study of human OSE exist. To characterize OSE cells in culture,
explants of ovarian surface from normal ovary of premenopausal women were grown on plastic, glass, and collagen gel in 25%
fetal bovine serum/Waymouth's medium 752/1. About 25% of explants produced epithelial outgrowths. Morphologically, these outgrowths
resembled OSE in vivo and endothelial and mesothelial cells in culture, but they differed from cultured ovarian stromal, granulosa,
and luteal cells. Only OSE among ovarian cell types were intensely keratin positive by immunofluorescence. Keratin also distinguished
OSE cells from the keratin-negative endothelial cells. Most but not all OSE colonies tested showed 17β-hydroxysteroid dehydrogenase
(HSD) activity, which was absent in peritoneal mesothelial cells. Colonies from most patients were limited to a few millimetres
and became stationary within a few weeks. Changes that accompanied cessation of growth included senescence, increased keratin
content, or the formation of multicellular papillary aggregates. With time, OSE cells tended to assume a fibroblast-like morphology
but remained keratin positive and continued to resemble OSE by scanning electron microscopy (SEM). Subcultured OSE cells persisted
in a stationary keratin-positive form for many weeks. Throughout this study, all pavementlike epithelial outgrowths that were
contiguous with an explant stained for keratin; thus, such colonies can be assumed to be OSE. Conversely, fibroblast-shaped
cells may represent OSE as indicated by keratin content and SEM appearance. The methods presented here permit culture of normal
human OSE under conditions in which the cells exhibit morphologic plasticity, variable 17β-HSD activity, and presence of keratin.
Supported by a grant and a research associateship to N. A. by the National Cancer Institute of Canada. 相似文献
14.
R. Niles K. C. Kim B. Hyman T. Christensen K. Wasano J. Brody 《In vitro cellular & developmental biology. Plant》1988,24(5):457-463
Summary Studies on the regulation of differentiation in airway epithelial cells have been hampered by the lack of cell culture systems
that differentiate in vitro. One such system that does exhibit differentiation is hamster tracheal epithelial cells (HTE).
A major problem with this system, however, is that at the time cells differentiate, they lyze the collagen gel upon which
they grow, resulting in termination of the culture. Here we report that by growing the HTE cells at 32° instead of 37°C we
can totally prevent lysis of the collagen gel. Cells grown at this lower temperature maintain their differentiated phenotype
as evidenced by abundant mucus granules and the secretion of authentic mucus glycoproteins into the culture media. We have
also developed a method for subculturing the primary cells which allows growth and differentiation in secondary culture. The
HTE cells were capable of being passaged at least three times and did not become transformed as judged by their inability
to grow in soft agar and to produce tumors in syngeneic animals. This improved HTE cell culture system will allow detailed
studies on the mechanisms which regulate growth, differentiation, and mucus secretion in surface airway epithelial cells.
This work was supported in part by grants HL-19717 and HL-36854 from the National Institutes of Health, Bethesda, MD. 相似文献
15.
16.
Anna L. Trifillis Myong Won Kahng 《In vitro cellular & developmental biology. Plant》1990,26(5):441-446
Summary We have developed an in vitro model of human papillary collecting duct cells isolated from cadaver kidneys using methods similar
to those we previously reported for the isolation of human proximal tubule cells. To date we have isolated papillary collecting
duct cells from 100 normal human kidneys. Papillae were dissected and digested in Cellgro containing 400 U/ml collagenase.
Cells were plated on fibronectin-coated culture flasks at a density of 104 live cells/ml in Cellgro supplemented with insulin and 10% fetal bovine serum. Confluent monolayers, which were able to withstand
600 mOsm for 8 h, were obtained within 10 to 15 d. Cells of primary isolates and first passages exhibited epithelial cell ultrastructure
including cell junctions, microvilli, and cilia. A dark-brown reaction product was observed in these cells when stained by
the immunoperoxidase method with peroxidase-labeled peanut lectin (Arachis hypogaea), which binds specifically to human distal tubule and collecting duct cells. These cells were negative for Factor-VIII (a
marker for endothelial cells) and γ-glutamyltransferase (a marker for proximal tubule cells). High activities of the glycolytic
enzyme pyruvate kinase and arginine vasopressin-stimulated cAMP production in these cells are consistent with a distal nephron
origin. The results indicate that human collecting duct cells can be isolated and cultured to provide an in vitro system to
probe pathogenetic mechanisms of potential nephrotoxins.
Part of this work was presented at a Symposium of the Center for Alternatives to Animal Testing, April 4–5, 1989, Johns Hopkins
Medical Institutions, Baltimore, MD 21205.
This work was supported in part by grants R01-AI24179, PO1-A804393 for the Public Health Service, U.S. Department of Health
and Human Services, and by a grant from the National Kidney Foundation, Baltimore, MD affiliate. 相似文献
17.
D. Kirk R. J. B. King Judy Heyes Linda Peachey P. J. Hirsch R. W. T. Taylor 《In vitro cellular & developmental biology. Plant》1978,14(8):651-662
Summary Separation of human endometrium into its epithelial and stromal components has been achieved through collagenase digestion
and has permitted a study of these two cell populations under specific experimental culture conditions. The stromal cell populations
showed a progesterone response, were easily handled in culture, and displayed a limited in vitro life span typical of human
diploid fibroblasts. In contrast, epithelium only survived in shortterm primary culture and showed no clear hormone response.
High-density epithelial cultures remained viable for longer periods in culture. Comparisons between resurfacing endometrial
epithelial cells in vivo and epithelial cells migrating from explants in vitro suggested that this initial epithelial migration
in vitro was the counterpart of the repair response in vivo.
We are much in debt to Dr. R. C. Hallowes (Department of Pathology, Imperial Cancer Research Fund) for his guidance and encouragement
throughout the course of this work. We also gratefully acknowledge Dr. P. N. Riddle (Time-Lapse Cinematography Unit, Imperial
Cancer Research Fund) for carrying out the time-lapse cinematography; Mrs. Lyn Rolph (Stereoscan Unit, Bedford College, University
of London) for assisting with the SEM; and Mr. G. D. Leach for his competent help with the photography. 相似文献
18.
Summary Growth characteristics of human esophageal epithelial cells have been determined in primary explant and serial culture. Normal
human esophagus was obtained from donor patients in a heart/lung transplantation program; tissue obtained at autopsy (6 to
22 h after death) was not viable. When mucosal specimens (1.5 mm2) were explanted on a plastic surface and attached with a plasma clot, 35% of explants detached from the surface within 48
h. The addition of epsilon amino caproic acid (EACA) to the culture medium increased explant attachment of 93% (P<0.001). Outgrowth kinetics were similar in both the presence and absence of EACA. No advantage of human serum over nonhuman
sera was observed in primary culture. Esophageal epithelium could be frozen in 10% dimethyl sulfoxide without affecting growth
kinetics. Addition of dexamethasone (DEX) significantly altered esophageal cell morphology in primary culture and increased
viability on serial culture. Studies of pH revealed an optimum at pH 7.4 with significantly decreased growth occuring at 6.8
and no growth at 6.2.
Esophageal cells in primary explant cultures could be released by trypsin and passaged two additional times with an eightfould
increase in total number. An increased rate of attachment and multiplication was observed for cells plated on a collagen substrate
compared to platic.
The addition of EACA and DEX to the culture media and the subculture on a collagen substrate provide a method for the isolation
and serial cultivation of human esophageal cells from biopsy-sized specimens of normal esophageal epithelium.
Supported in part by Grant AM—14121 of the United States Public Health Service. A preliminary report of this work appeared
in Clin. Res. 30: 93A; 1982. 相似文献
19.
Dr. Werner Müller-Glauser 《Cell and tissue research》1981,221(1):147-156
Summary In previous studies the differentiation of the epithelium in the human hard palate has been described stereologically using parameters expressed per unit tissue volume. Since single epithelial cells represent the true biological units of this tissue, it became necessary to estimate the absolute size of such cells in order to transform density data into absolute data. Therefore, in the present study, a stereological method (originally developed for myocyte volume determination) was tested in terms of its applicability to stratified epithelia; the absolute size of differentiating epithelial cells was determined in the epithelium of the human hard palate. The results suggest that (1) rather precise determination of epithelial cell size is possible by using the modified myocyte volume determination, and (2) the average cell volumes are 926 ± 148, 4,111 ± 1,619, 4,394 ± 551 m3 for the stratum basale, the upper stratum spinosum and the stratum granulosum, respectively. The results are discussed with respect to methodology and to differentiation phenomena in the epithelium of the human hard palate. 相似文献
20.
Shahnaz Hashmi Dairkee Carlene M. Blayney David M. Asarnow Helene S. Smith Adeline J. Hackett 《In vitro cellular & developmental biology. Plant》1985,21(6):321-327
Summary The intermediate filaments of most epithelial cells in vivo consist solely of cytokeratins. Using monoclonal antibodies to
vimentin or keratin, we have examined the expression of vimentin in homologous specimens of frozen tissue sections and primary
cultures of normal human mammary epithelium. In frozen sections, only epithelial cells reacted with the antikeratin antibody,
whereas antivimentin reactivity was associated with stromal cells. All epithelial cultures were positive for cytokeratin and
in addition coexpressed vimentin as strongly as cultured fibroblasts and as early as the 4th d after initiation of the culture.
Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis of cytoskeletal preparations
of secondary cultures of normal mammary epithelium have also demonstrated the appearance of a moiety identical to the vimentin
found in cultured fibroblasts. Our observations are consistent with the hypothesis that vimentin expression is induced, possibly
as a result of changes in cell shape or growth rate, when cells are freed from three-dimensional restirctions imposed by the
tissue of origin 相似文献