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1.
Gerardo Martínez-López L. M. Black 《In vitro cellular & developmental biology. Plant》1977,13(11):777-784
Summary The optimal composition of a medium for tissue culture of cells from the leafhopperAgallia constricta was estimated from experiments in which the rate of growth of the cells was measured at different concentrations of each
component. Wound tumor virus and theconstricta variety of potato yellow dwarf virus multiply, inA. constricta when these viruses are transmitted from one plant to another. Differences weredetected in the suitablility of different batches
of fetal bovine serum (after heating at 56°C for 30 min), and histidine as components in the tissue culture medium. Without
heat treatment even thebest fetal bovine serum was not suitable. Estimated optimal concentrations of fetal bovine serum, histidine,
salts, dextrose, lactalbumin hydrolysate and yeast autolysate were determined. Fungizone (amphotericin B) at 50 or 100 mg
per 1 caused harmful effects; effective concentrations of penicillin, neomycin and steptomycin did not. Combinations of histidine-HCl
and histidine (free-base) made it possible to prepare buffered medium at my pH between 6.0 and 7.0. Optimal growth ofA. constricta cells occurred at pH 6.43, and ofAceratagallia sanguinolenta at pH 6.30. Osmotic pressures of the new medium between 360 and 405 mOSM were better than lower osmotic pressures. The new
medium was still suitable for growth of the cells after, storage for 6 months at 4°C.
Portion of a thesis submitted for the Ph.D. degree by the senior author to the Graduate College of the University of Illinois.
This research was supported in part by Grant GB 20915 from the National Science Foundation and Grant AI 6392 from the National
Institutes of Health. 相似文献
2.
Eileen A. Friedman Paul J. Higgins Martin Lipkin Hiromi Shinya Alvin M. Gelb 《In vitro cellular & developmental biology. Plant》1981,17(7):632-644
Summary Human colonic epithelial cells from three classes of benign tumors have been reproducibly cultured free of fibroblasts for
8 wk using a supplemented Medium 199 (M 199S). The cultured colonic cells were identified as epithelial by the presence of
junctional complexes (tight junctions, gap junctions, and desmosomes), a brush border on the apical surface, keratin fibrils,
and by both a close-packed columnar or cuboidal morphology and the capability to transport water and ions to form hemicysts.
Colony formation was initiated by groups of epithelial cells, not by single cells, and was inhibited by cocultivation with
either lethally irradiated 3T3 cells or human diploid fibroblasts. Enhancement of epithelial colony formation was observed
following culture on nonadherent, “floating” substrates compared with substrates attached directly to the bottom of the culture
dish.
Replication of epithelial cells in M 199S from the class of benign colonic tumors least prone to malignancy, the tubular,
was significantly enhanced by epidermal growth factor (EGF). In contrast, EGF did not stimulate the growth of cells in M 199S
from the other classes of benign tumors, the villotubular and the villous, which exhibit more malignant potential. These data
imply that premalignant colonic epithelial cells lose responsiveness to growth modulation by EGF as they progress toward frank
carcinoma.
This study was supported by NCI Contract N01-CP43366 to M. L. and NCI Grant 1-R26-CA 28822 to E. F. 相似文献
3.
Peter I. Lelkes Esther Ramos Victor V. Nikolaychik Dawn M. Wankowski Brian R. Unsworth Thomas J. Goodwin 《In vitro cellular & developmental biology. Animal》1997,33(5):344-351
Summary The aim of this study was to test the versatility of a new basal cell culture medium, GTSF-2. In addition to traditional growth-factors,
GTSF-2 contains a blend of three sugars (glucose, galactose, and fructose) at their physiological levels. For these studies,
we isolated normal endothelial cells from human, bovine, and rat large blood vessels and microvessels. In addition, GTSF-2
was also tested as a replacement for high-glucose-containing medium for PC12 pheochromocytoma cells and for other, transformed
cell lines. The cell growth characteristics were assessed with a novel cell viability and proliferation assay, which is based
on the bioreduction of the fluorescent dye, Alamar Blue. After appropriate calibration, the Alamar Blue assay was found to
be equivalent to established cell proliferation assays. Alamar Blue offers the advantage that cell proliferation can be measured
in the same wells over an extended period of time. For some of the cell types (e.g., endothelial cells isolated from the bovine
aorta, the rat adrenal medulla, or the transformed cells), proliferation in unmodified GTSF-2 was equivalent to that in the
original culture media. For others cell types (e.g., human umbilical vein endothelial cells and PC12 cells), GTSF-2 proved
to be a superior growth medium, when supplemented with simple additives, such as endothelial cell growth supplement (bFGF)
or horse serum. Our results suggest that GTSF-2 is a versatile basal medium that will be useful for studying organ-specific
differentiation in heterotypic coculture studies. 相似文献
4.
Katrin Engelmann Peter Friedl 《In vitro cellular & developmental biology. Plant》1989,25(11):1065-1072
Summary Long-term cultivation of human corneal endothelial cells (HCEC) was optimized with respect to different components of the
culture system: 25 different nutrient media, different sera, 6 mitogens and various substrates were tested in their ability
to influence clonal growth and morphology of HCEC. F99, a 1∶1 mixture of the two media M199 and Ham’s F12, was the most effective
basal medium in promoting clonal growth of HCEC. Among various sera, human serum and fetal bovine serum showed optimal growth
promoting activities in combination with F99, whereas newborn bovine serum (NBS) was by far superior for the development of
a typically corneal endothelial morphology. Crude fibroblast growth factor (FGF), or alternatively endothelial cell growth
supplement, was absolutely essential for clonal growth of HCEC at low serum concentrations, for example 5% NBS. Formation
of a monolayer with a morphology similar to corneal endothelium in vivo was observed only on culture dishes coated with basal
membrane components such as collagen type IV, laminin, or fibronectin. The most pronounced effect on morphologic appearance
was obtained by culturing the cells on the extracellular matrix (ECM) produced by bovine corneal endothelial cells. Moreover,
ECM could substitute for crude FGF in clonal growth assays. 相似文献
5.
A method for the isolation and serial propagation of keratinocytes,endothelial cells,and fibroblasts from a single punch biopsy of human skin 总被引:5,自引:0,他引:5
Summary When multiple types of cells from normal and diseased human skin are required, techniques to isolate cells from small skin
biopsies would facilitate experimental studies. The purpose of this investigation was to develop a method for the isolation
and propagation of three major cell types (keratinocytes, microvascular endothelial cells, and fibroblasts) from a 4-mm punch
biopsy of human skin.
To isolate and propagate keratinocytes from a punch biopsy, the epidermis was separated from the dermis by treatment with
dispase. Keratinocytes were dissociated from the epidermis by trypsin and plated on a collagen-coated tissue culture petri
dish. A combination of two commercial media (Serum-Free Medium and Medium 154) provided optimal growth conditions.
To isolate and propagate microvascular endothelial cells from the dermis, cells were released following dispase incubation
and plated on a gelatin-coated tissue culture dish. Supplementation of a standard growth medium with a medium conditioned
by mouse 3T3 cells was required for the establishment and growth of these cells. Epithelioid endothelial cells were separated
from spindle-shaped endothelial cells and from dendritic cells by selective attachment toUlex europeus agglutinin I-coated paramagnetic beads.
To establish fibroblasts, dermal explants depleted of keratinocytes and endothelial cells were attached to plastic by centrifugation,
and fibroblasts were obtained by explant culture and grown in Dulbecco’s modified Eagle’s medium (DMEM) containing fetal bovine
serum (FBS).
Using these isolation methods and growth conditions, two confluent T-75 flasks of keratinocytes, one confluent T-25 flask
of purified endothelial cells, and one confluent T-25 flask of fibroblasts could be routinely obtained from a 4-mm punch biopsy
of human skin. This method should prove useful in studies of human skin where three cell types must be grown in sufficient
quantities for molecular and biochemical analysis. 相似文献
6.
Routine culturing of normal,dysplastic and malignant human mammary epithelial cells from small tissue samples 总被引:5,自引:0,他引:5
Joanne T. Emerman Darcy A. Wilkinson 《In vitro cellular & developmental biology. Plant》1990,26(12):1186-1194
Summary We compared the growth and morphology of normal, dysplastic and malignant human mammary epithelial cells (HMEC) in medium
containing 5% human serum, a serum-free medium (32) and serum-free medium with a low Ca++ concentration. Tissues were dissociated and epithelial organoids or single cells were seeded onto collagen-coated dishes.
The cells grew in serum-containing medium, but growth of fibroblasts was also stimulated. The serum-free medium consistently
selected for and stimulated the growth of epithelial cells. There was little advantage in reducing the Ca++ concentration to further increase cell yield. This serum-free primary culture system allows us to routinely prouce sufficient
numbers of HMEC from small tissue samples for molecular biological investigations. Furthermore, the maintenance of cells in
a defined medium can provide a system for evaluating the direct effects of factors on gene expression.
This work was supported by a grant from the National Cancer Institute of Canada and funds contributed by Mr. B. T. Wharton
in memory of his wife, Nadia. 相似文献
7.
Zea Borok Spencer I. Danto Stephanie M. Zabski Edward D. Crandall 《In vitro cellular & developmental biology. Animal》1994,30(2):99-104
Summary Isolated type II pneumocytes grown in serum on tissue culture-treated polycarbonate filters form monolayers with characteristic
bioelectric properties, and change morphologically with time in culture to resemble type I cells. Concurrently, the cells
express type I cell surface epitopes, making this a potentially useful in vitro model with which to study regulation of alveolar
epithelial cell function and differentiation. To define specific soluble growth factors and matrix substances that may regulate
these processes, it would be preferable to culture isolated pneumocytes de novo under completely defined, serum-free conditions.
In this study, we developed a completely defined serum-free medium that is capable of supporting alveolar epithelial cells
in primary culture, allowing the formation of monolayers with characteristic bioelectric and phenotypic properties. Freshly
isolated rat type II cells were resuspended in completely defined serum-free medium and plated de novo on polycarbonate filters.
Plating efficiency, bioelectric properties, morphology, and binding of a type I cell-specific monoclonal antibody were determined
as functions of time. Plating efficiency plateaus at about 14% by Day 3 in culture. Transepithelial resistance rises to high
levels, peaking at 1.76±0.14 KΩ-cm2 by Day 5 in culture. Short-circuit current peaks on Day 3 in culture at 2.71±0.35 μA/cm2. With time, the cells gradually become flattened with protuberant nuclei and long cytoplasmic extensions, more closely resembling
type I cells, and begin to express a type I cell surface epitope. These observations indicate that it is feasible to culture
alveolar epithelial cell monolayers under completely defined serum-free conditions de novo. This culture system should prove
useful for identifying soluble growth factors and matrix substances that modulate alveolar epithelial cell biological properties. 相似文献
8.
Elena Torreggiani Marika Rossini Ilaria Bononi Silvia Pietrobon Elisa Mazzoni Maria Rosa Iaquinta Carlo Feo John Charles Rotondo Paola Rizzo Mauro Tognon Fernanda Martini 《Journal of cellular physiology》2019,234(7):9895-9905
Procedures for in vitro culturing of human primary keratinocytes from normal colon mucosa specimens have not been fully feasible, thus far. The protocol described herein allows primary keratinocytes from small tissue fragments of colorectal mucosa biopsies to grow in vitro. The procedure develops in three steps: (a) the enzymatic digestion of the tissue biopsy; (b) the use of cloning rings to purify primary keratinocyte colonies, (c) a defined keratinocyte medium to grow these cells in long-term culture. Our cultural method enables normal primary keratinocytes to be obtained by simple and rapid techniques. In our culture condition, primary keratinocytes express specific epithelial markers. Colorectal mucosa keratinocyte colonies require approximately 2 weeks to grow. Compared with previous approaches, our protocol provides a valuable model of study for human primary keratinocytes from normal colorectal (NCR) mucosa both at the cellular and molecular levels. It is well known, that different mutations occurring during the multistep process of carcinogenesis in the NCR mucosa, are strictly associated to the onset/progression of the colorectal carcinoma. On this ground, normal keratinocytes grown with our protocol, may represent an innovative tool to investigate the mechanisms that lead to colorectal carcinoma and other diseases. Our innovative procedure may allow to perform comparative investigations between normal and pathological colorectal cells.
9.
The 3-dimensional culture of human tumor spheroids under standardized medium conditions may reveal information on specific biological parameters that could be masked in serum-supplemented media. Spheroids derived from human tumor cells are growth retarded in media free of serum. Ex-Cyte IV is a substance derived from human blood that can be used to improve growth in tissue culture. In this study the growth of spheroids from four different human tumor cell lines was studied when grown in medium free of serum, medium supplemented with varying concentrations Ex-Cyte IV, and medium supplemented with foetal calf serum (FCS). The parameters used for comparisons were growth rate, growth enhancement, clonogenicity and cell cycle distribution.The four cell lines showed different growth rates in serum-free medium, which were increased to different extents when Ex-Cyte IV or FCS were added. The growth enhancing effect induced by Ex-Cyte IV was differently concentration dependent for each cell line. The clonogenicity of cells grown as spheroids in serum-free medium was lower than in spheroids grown in supplemented media. There was no difference in clonogenicity between the differently supplemented media. All four cell lines responded to growth in serum-free medium with a drop in the S-phase and G2M phase.The present study provides a novel approach to the study of human tumor cells in 3-dimensional culture under defined conditions. The human serum derived substance Ex-Cyte IV may provide a method to obtain information on specific biological parameters that could be masked in serum-supplemented media. 相似文献
10.
Chongtao Zhu Jiankun Liu Bin He Xiaowen Qu Daizhi Peng 《Journal of cellular biochemistry》2019,120(8):12182-12191
In this study, we aimed to investigate the phenotypic characteristics of human immortal skin keratinocytes (HaCaT) cells and the role of acellular dermal matrix (ADM) in coculture system of HaCaT cells and ADM. Flow cytometry was used to examine the cluster of differentiation (CD) makers of HaCaT cells. Apoptosis analysis was applied to detect the apoptosis rate of HaCaT cells. Morphological observation of ADM isolated from the reticular layer of Sprague-Dawley rat dermis was utilized to evaluate the morphological structure of ADM. Methylthiazolyl tetrazolium (MTT) assay and morphological experiments were further used to confirm the scaffold role of ADM in HaCaT cells. A wound-healing mice model accompanied by HaCaT-ADM scaffold transplantation was performed to further verify the function of HaCaT-ADM scaffold. Our results showed that CD71, CD49f, K19, and CD29 were highly expressed in HaCaT cells, and the percentage of apoptosis cells was significantly increased, which represented that HaCaT cells had much stronger capacities of adhesion and proliferation than normal human keratinocytes. Additionally, the morphological structure of ADM presented many natural microbores, which made cells rapidly grow on ADM. The results exhibited that the HaCaT cells indeed promptly proliferate on ADM and easily grow into the microbores of ADM. Finally, an in vivo experiment further confirmed that the transplantation of the HaCaT-ADM scaffold into the dorsal skin of a wound-healing mice model could gradually repair the injured wound. Thus, these findings indicated that HaCaT cells might be as seed cells to develop skin tissue engineering and the HaCaT-ADM scaffold might be a better candidate to promote skin repair and regeneration. 相似文献
11.
CHO工程细胞 (11G-S) 悬浮培养的无血清培养基的设计 总被引:1,自引:1,他引:1
以悬浮适应的表达重组尿激酶原 (Pro-urokinase,pro-UK) CHO工程细胞系11G-S为对象,采用Plackett-Burman实验设计及响应面分析法,设计支持CHO工程细胞 (11G-S) 悬浮生长的无血清培养基。以细胞密度为评价指标,在单因素实验的基础上采用Plackett-Burman实验设计对影响细胞生长的培养基添加成分进行考察,确定了3种对细胞生长明显促进作用的培养基添加成分:胰岛素、转铁蛋白及腐胺。继而利用响应面法分析了这3种添加成分的最佳水平范围,设计了一种适用于CHO工程细胞 (11G-S) 悬浮培养的无血清培养基SFM-CHO-S。11G-S细胞在SFM-CHO-S批次悬浮培养的细胞最大生长密度达到4.12×106 cells/mL,pro-UK的最大累积活性达到5 614 IU/mL,培养效果优于商品化的同类无血清培养基。 相似文献
12.
Growth of normal human mammary cells in culture 总被引:27,自引:0,他引:27
M. Stampfer R. C. Hallowes A. J. Hackett 《In vitro cellular & developmental biology. Plant》1980,16(5):415-425
Summary Reduction mammoplasty tissue was used to obtain short-term cultures of human epithelial cell populations. Digestion of tissue
with collagenase and hyaluronidase resulted in cell clusters (organoids) resembling ductal and alveolar structures; these
could be separated by filtration from the stromal components. Epithelial outgrowth from these organoids was greatly enhanced
by the addition of conditioned medium from other human epithelial and myoepithelial cell lines. Additionally, the mammary
epithelial growth was stimulated by insulin, hydrocortisone, epidermal growth factor, and steroid hormones. With this enriched
nutritional environment, active cell division could be maintained for 1 to 3 months and cells could be serially subcultured
1 to 4 times.
This research was supported by Grant PDT-72 from the American Cancer Society and Grant CP-70510 from the National Institutes
of Health. 相似文献
13.
Martha R. Stampfer 《In vitro cellular & developmental biology. Plant》1982,18(6):531-537
Summary Addition of cholera toxin to human mammary epithelial cultures derived from reduction mammoplasties and primary carcinomas
greatly stimulated cell growth and increased the number of times the cells could be successfully subcultured. Other agents
known to increase intracellular cAMP levels were also growth stimulatory. The increased growth potential conferred by cholera
toxin enhances the usefulness of this cell culture system.
This work was supported by USPHS Grant CA-24844 from the National Cancer Institute and Grant CD-61B from the American Cancer
Society. 相似文献
14.
Isolation and culture of airway epithelial cells from chronically infected human lungs 总被引:5,自引:0,他引:5
Scott H. Randell Diana L. Walstad Ute E. Schwab Barbara R. Grubb James R. Yankaskas 《In vitro cellular & developmental biology. Animal》2001,37(8):480-489
We describe procedures for isolating and culturing airway epithelial cells from chronically infected human lungs. Experience in our laboratory demonstrated the need to balance pathogen eradication against antibiotic toxicity to epithelial cells. To provide a logical basis for antibiotic selection and dose, we systematically analyzed the cytotoxicity of antibiotics useful against typical pathogens. Alone, colistin, ciprofloxacin, doxycycline, and tobramycin were moderately toxic at concentrations close to those used in cell culture, whereas amphotericin, ceftazidime, chloramphenicol, imipenem, meropenem, piperacillin, sulfamethoxazole/trimethoprim, and vancomycin were nontoxic even at concentrations many times the antimicrobial level. Epithelial cytotoxicity of combined antibiotics was additive, with no evidence of competition or synergism. Antibiotics had little effect on initial cell attachment and did not acutely lyse cells, but inhibited subsequent growth. Interestingly, cytotoxicity decreased markedly with increasing epithelial cell density. Cystic fibrosis (CF) and non-CF epithelial cells showed no differences in sensitivity to the antibiotics tested and initial exposure to antibiotics did not affect the electrophysiologic properties of resistance or short circuit current in well-differentiated cells. Tailored combinations of antibiotics at appropriate doses killed even multidrug-resistant bacteria. Thus, epithelial cells can usually be cultured from chronically infected CF airways. 相似文献
15.
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
16.
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
17.
A new diploid nontumorigenic human breast epithelial cell line isolated and propagated in chemically defined medium 总被引:10,自引:0,他引:10
P. Briand O. W. Petersen B. Van Deurs 《In vitro cellular & developmental biology. Plant》1987,23(3):181-188
Summary A new, nontumorigenic human breast epithelial cell line, HMT-3522, has been established from fibrocystic breast tissue. Cells
were explanted and propagated in chemically defined medium including insulin, transferrin, epidermal growth factor, hydrocortisone,
estradiol, prolactin, and Na-selenite. The epithelial nature of the cell line was established by immunocytochemical detection
of cytokeratins. Moreover, electronmicroscopy revealed monolayers of polarized cells connected by desmosomes and provided
with apical microvilli. Milk fat globule membrene antigen, specific for the apical membrane domain of normal, luminal breast
epithelial cells, was expressed only in confluent cultures where some cells overlaid others, indicating “stem cell”-like properties.
After 25 to 30 passages, the cells are diploid with a few marker chromosomes and loss of chromosomes in the D-group. The cells
are nontumorigenic in athymic mice; they lack estrogen receptors, and estradiol does not stimulate growth. The HMT-3522 cell
line may represent a useful model for the study of brest cell differentiation and carcinogenesis in vitro.
This work was supported by a grant from the Danish Cancer Society. 相似文献
18.
The effects of several different substances, including insulin, transferrin, ethanolamine, selenite and butyrate on the growth of murine hybridoma 2F7 cells, which secrete monoclonal antibody against small cell lung cancer, were investigated, and a serum-free medium SFMI was formulated. The effects of taurine, spermidine, progesterone and adenine on the cell growth were tested further on the basis of the medium SFMI, and a modified serum-free medium SFM II was established. On the basis of medium SFM II, the substitution tests of ferric citrate for transferrin were carried out, and it was found that transferrin could be replaced. The experiments suggested that the formulated serum-free medium was suitable for 2F7 cell growth and monoclonal antibody secretion, and thus facilitated subsequent purification of monoclonal antibody.Abbreviations BSA
bovine serum albumin
- CS
calf serum
- DMEM
Dulbecco's modified Eagle's medium
- ELISA
enzyme-linked immunosorbant assay
- McAb
monoclonal antibody
- PEG
polyethylene glycol
- SFM
serum-free medium 相似文献
19.
R Benali F Dupuit M Chevillard J Jacquot B Haye E Puchelle 《Biology of the cell / under the auspices of the European Cell Biology Organization》1991,73(1):49-56
Bovine tracheal gland (BTG) cells in culture show an epithelial-fibroblastoid transition after several passages. To investigate these BTG cell phenotype changes, we studied the effects of both the culture medium and passage number on the expression of epithelial cytoskeletal proteins and glandular serous cell markers. We also analyzed the intracellular cAMP level in the basal state and after adrenergic stimulation. Three culture media were used: 1) serum-free defined medium (SFDM); 2) medium supplemented with 2% Ultroser G; and 3) medium supplemented with 10% fetal calf serum (FCS). Using immunofluorescence microscopy, we showed that, in the first 4 passages whatever the culture conditions, BTG cells expressed immunoreactivities to cytokeratin filaments and desmoplakins I and II, whereas vimentin filaments were not detected. After four passages, BTG cells cultured in 10% FCS or 2% Ultroser G became progressively fibroblastoid and showed immunoreactivities to both vimentin and cytokeratin intermediate filaments. No immunoreactivity to vimentin filaments was observed on BTG cells cultured in a SFDM. Using biochemical analysis, we showed that basal levels of cAMP in cultured BTG cells and lysozyme secretion by these cells vary according to the culture medium and passage number. It was higher in BTG cells cultured in a SFDM compared to that recovered from cells cultured in medium supplemented with Ultroser G or FCS. Whatever the culture medium, BTG cells responded to stimulation by isoproterenol. However, the results of stimulation in a SFDM were higher than in Ultroser G or FCS supplemented medium. We conclude that the BTG epithelial cell organization and the regulation of biosynthesis of secretory proteins by these cells in culture depend on both the culture medium and passage number.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
20.
Radu E Simionescu O Regalia T Dumitrescu D Popescu LM 《Journal of cellular and molecular medicine》2002,6(4):593-598
Epidermal stem cells (ESC) are responsible for maintaining skin cellular homeostasis, as they give rise to fast-dividing transit amplifying cells committed to terminal differentiation, while retaining their self-renewal capacity. However, no pure ESC cultures are available and no highly specific cytochemical marker was identified. We report here the experimental conditions allowing the selective enrichment in ESC, using cultured adult human keratinocytes. The main step was the selection of cells able to rapidly adhere to human collagen type IV in vitro . Thus, an increased proportion of putative ESC of about 65% was obtained, as demonstrated by p63 expression. 相似文献