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1.
Jill M. Siegfried Karen G. Nelson Jane L. Martin David G. Kaufman 《In vitro cellular & developmental biology. Plant》1984,20(1):25-32
Summary Histochemical techniques have been applied to the identification of cell types cultured from human endometrium. Previous work
from this laboratory characterized two principtal cell types found in cultures of endometrium: a mature epithelial cell and
another cell which was classified as the endometrial stromal cell based on light and electron microscopy. In this report we
compare the histochemical staining of endometrial tissue in frozen sections to that of cultured cells. These results confirm
the epithelial and stromal nature of the respective cell types. Several markers were found that could distinguish between
cells of epithelial and stromal origin. The enzymes alkaline phosphatase, γ-glutamyltranspeptidase, peroxidase, and β-glucuronidase
were localized in glandular and surface epithelia in frozen sections and in colonies of epithelial cells in culture. Stroma
in frozen sections and cultured stromal cells contained leucine aminopeptidase and fibronectin. Epithelial sections and in
culture could also be distinguished from cells of stromal origin by preferential binding of lotus and peanut lectin. Several
other markers were found in both endometrial epithelium and stroma.
J. M. S. was recipient of National Research Service Award CA09156 (National Cancer Institute); K. G. N. was recipient of National
Research Service Award ES07017 (National Institute of Environmental Health Sciences); and D. G. K. was recipient of Research
Career Development Award CA00431 from the National Cancer Institute, Bethesda, MD. Supported by Grant CA 31733 from the National
Cancer Institute, Bethesda, MD. 相似文献
2.
Dianne M. Rausch Sidney B. Simpson Jr. 《In vitro cellular & developmental biology. Plant》1988,24(3):217-222
Summary The immune suppressed lizard,Anolis carolinensis, can be used to test for in vivo tumor production by cell lines derived from a variety of ectothermic vertebrates. Cell lines
tested for tumor production were also assessed for loss of attachment-dependent proliferation and contact inhibition of cell
overlap. The results demonstrate that the criteria standardly used to assess transformation and neoplastic change in cultured
mammalian cells apply equally well to cultured cells from ectotherms.
Supported by grants AG01476 and NS24162 from the National Institutes of Health, Bethesda, MD. 相似文献
3.
Summary The interactions of vascular smooth muscle cells with growth modulators and extracellular matrix molecules may play a role
in the proliferation and migration of these cells after vascular injury and during the development of atherosclerosis. Time-lapse
cinematographic techniques have been used to study cell division and migration of bovine carotid artery smooth muscle cells
in response to matrix molecules consisting of solubilized basement membrane (Matrigel) and type I collagen. When cells were
grown adjacent to Matrigel, both migration and cell proliferation were increased and interdivision time was shortened. Cells
grown in Matrigel or in type I collagen had markedly reduced migration rates but interdivision time was not altered. Further,
diffusible components of the Matrigel were found to stimulate proliferation of the smooth muscle cells.
This work was supported by grants HL35684 and SCOR HL14212 from the National Institutes of Health, Bethesda, MD. 相似文献
4.
Warren I. Schaeffer Betsy F. von Kreuter Elizabeth A. Reichard Gordon H. Sato 《In vitro cellular & developmental biology. Plant》1983,19(2):108-110
Summary A hormonally defined medium consisting of a 1∶1 mixture of Dulbecco's modified Eagle's medium and nutrient mixture F12 supplemented
with 50 μg/ml each of insulin and transferrin was found to grow late passage malignantly transformed cells of the RL-PR-C
rat liver cell culture. The same medium formulation would not grow the early passage, normal, diploid counterpart of the RL-PR-C
cell culture. When mixtures of the early and late passage cells were made, only the late passage cells would grow, thus providing
a selection system for the late passage cells.
This work was supported by Grant PHS SO7 05429-20-3 from the National Institutes of Health, Bethesda, MD. 相似文献
5.
Jerry Y. Niederkorn Dale R. Meyer John E. Ubelaker James H. Martin 《In vitro cellular & developmental biology. Plant》1990,26(9):923-930
Summary A widely utilized rabbit corneal cell line, SIRC, was characterized ultrastructurally and immunohistologically. Although SIRC
cells are often described as being of epithelial origin, important ultrastructural and antigenic characteristics indicate
that these cells are fibroblastic and not epithelial. SIRC cells lack desmosomes, cytoplasmic filaments, and cytokeratin—structures
that are characteristic of corneal epithelial cells. By contrast, the dendritic morphology, presence of vimentin, and the
extensive dense accumulations of ribosomes and rough endoplasmic reticulum are consistent with a fibroblastic phenotype. Collectively,
the morphology, ultrastructural features, and antigenic composition favor the hypothesis that SIRC cells are fibroblastic
cells (keratocytes) and not corneal epithelial cells.
This work supported in part by grant EY 07641 from the National Institutes of Health, Bethesda, MD, and an unrestricted grant
from Research to Prevent Blindness, Inc., New York. 相似文献
6.
A cloned rat thymic epithelial cell line established from serum-free selective culture 总被引:6,自引:0,他引:6
Arthur Piltch Paul Naylor Jun Hayashi 《In vitro cellular & developmental biology. Plant》1988,24(4):289-293
Summary A serum-free system has been developed for selective growth and long-term culture of rat thymic epithelial cells. The growth
media is a modification of McKeehan's WAJC 404, plus insulin, cholera toxin, dexamethasone, and epidermal growth factor. Cultures
have been continuously passaged and maintained for over 6 mo., and a cloned cell line, TEA3A1, has been established. These
cells are epithelial, judging by morphology and ultrastructure, and are positive for A2B5 and thymosin α markers for thymic
endocrine cells.
This work was partly supported by grant PCM-834 0582 from the National Science Foundation, Washington, DC, and grant P01 CA
37589-2 from the National Cancer Institute, Bethesda, MD. 相似文献
7.
Mark A. Hadley Stephen W. Byers Carlos A. Suárez-Quian daniel Djakiew Martin Dym 《In vitro cellular & developmental biology. Plant》1988,24(6):550-557
Summary Primary cultures of Sertoli cells maintained in conventional cultures on plastic culture vessels do not retain many of the
structural and functional properties of their in vivo counterparts. Sertoli cell phenotype is better maintained by incorporating
certain environmental parameters, intrinsic to the testic, into the Sertoli cell culture system. These environmental parameters
include a) high cell density, b) a unique extracellular matrix, c) a semipermeable support between the basal plasma membrane
of the cells and blood-derived nutrients in the interstitium, d) chemically distinct microenvironments at the apical and basal
surfaces of the cells, and e) cell-to-cell interactions among Sertoli cells and other testicular cell types. Using three variations
of Sertoli cell culture we have demonstrated the importance of each of these environmental parameters in obtaining a better
Sertoli cell culture model.
Paper presented a the 38th Annual Meeting of the Tissue Culture Association in Arlington, Virginia, in May 1987. The session
was chaired by Dr. Carlton H. Nadolney, member of the TCA Committee on Toxicity, Carcinogenesis and Mutagenesis Evaluation.
This work was supported by grant HD-16260 from the National Institutes of Health, Bethesda, MD, and a grant from the Mellon
Foundation. 相似文献
8.
Franklin Greif Harry S. Soroff R. Woodrow Setzer Lorne B. Taichman 《In vitro cellular & developmental biology. Plant》1988,24(10):985-989
Summary Epidermal keratinocytes grow in culture to form a stratified squamous epithelium. These cultures contain a replicating as
well as a terminally differentiating population and undergo surface desquamation. Epidermal growth factor (EGF) and cholera
toxin are usually employed as growth-promoting agents because they reduce the population doubling time; that is, the period
required to increase the total cell number twofold. There are three ways in which this reduction in population doubling time
could be achieved: (a) the time for one cell cycle or the cell cycle length may be shortened; (b) the number of cells that
withdraw from the cell cycle and terminally differentiate may be reduced; or (c) the number of cells that desquamate into
the medium over a set period of time may be reduced. We have explored these possibilities in growing cultures of epidermal
keratinocytes using a newly developed double-label assay. This assay gives a measure of both cell length and cell cycle withdrawal.
Results show that the growth enhancement induced by EGF and cholera toxin can be attributed primarily to a reduction in cell
cycle withdrawal and, to a lesser degree, to a reduction in cell cycle length. EGF and cholera toxin have no significant effect
on the rate of desquamation. A linear correlation was noted between cell cycle lengths and withdrawal, suggesting an interconnection
between the rate of cell renewal and the likelihood of undergoing terminal differentiation.
This research was supported by grant DE04511 from the National Institute of Dental Research, Bethesda, MD, and gifts from
the University Hospital Auxilliary, Health Sciences Center, SUNY Stony Brook, and the Suffolk County Volunteer Firefighter
Fund. 相似文献
9.
Summary A monolayer culture system has recently been developed for the extended growth and serial passage of normal rat mammary epithelial
(RME) cells. In this system the cells undergo greater than 20 population doublings when grown on type I collagen-coated tissue
culture dishes in Ham's F12 medium supplemented with insulin, hydrocortisone, epidermal growth factor, prolactin, progesterone,
cholera toxin, and 5% fetal bovine serum (FBS). The purpose of the present studies was to define additional growth factors
that would allow equivalent RME cell proliferation in serum-free medium. Ethanolamine (EA) was effective at reducing the FBS
requirements for RME cell proliferation and at its optimum concentration did so by greater than 20-fold. Even with optimum
levels of EA there was essentially no cell proliferation in the absence of FBS. However, addition of bovine serum albumin
(BSA) to the hormone, growth factor, and EA-supplemented medium resulted in substantial proliferation in the absence of serum,
and the further addition of transferrin (T) potentiated this effect. Thus, in this culture system, replacement of FBS with
EA, BSA, and T resulted in RME cell proliferation in primary culture which was equivalent to that obtained in the 5% FBS-containing
medium.
This work was supported by grant RR-05529 from the Division of Research Resources, National Institutes of Health, Bethesda,
MD, and by Public Health Service grant CA40064-01 from the National Cancer Institute, Bethesda, MD. 相似文献
10.
Longitudinal growth of skeletal myotubes in vitro in a new horizontal mechanical cell stimulator 总被引:7,自引:0,他引:7
Herman H. Vandenburgh Patricia Karlisch 《In vitro cellular & developmental biology. Plant》1989,25(7):607-616
Summary A new computerized mechanical cell stimulator device for tissue cultured cells is described which maintains the cells in a
horizontal position during mechanical stretching of up to 400% in substratum length. Mechanical stimulation of myogenic cells
in this device initiates several aspects of in vivo skeletal muscle organogenesis not seen in normal static tissue culture
environments. Embryonic skeletal muscle cells from avian m. pectoralis are grown in the device attached to the collagen-coated
elastic substratum. Dynamic stretching of the substratum in one direction for 3 d at a rate (0.35 mm/h) that simulates in
vivo bone elongation during development causes the myoblasts to fuse into parallel arrays of myotubes which are 2 to 4 times
longer than myotubes grown under static culture conditions. This longitudinal myotube growth is accompanied by increased rates
of cell proliferation and myoblast fusion. Prestretching the collagen-coated substratum before cell plating also results in
increased cell proliferation, myotube orientation, and longitudinal myotube growth. The effects of substratum stretching on
myogenesis in this model system thus occur by alterations in the cell’s extracellular matrix and not by acting directly on
the cells.
This work was supported by grant AR36266 from the National Institutes of Health, Bethesda, MD, and research grnat NAG2-414
from the National Aeronautics and Space Administration, Washington, DC. Parts of this work have appeard in abstract form,
J. Cell. Biochem. 12C:360; 1988. 相似文献
11.
David M. Halton Ward D. Peterson Bharati Hukku 《In vitro cellular & developmental biology. Plant》1983,19(1):16-24
Summary Procedures that involve cell cultures require careful quality control to avoid inter- and intraspecies contamination. We have
developed and electrophoresis technique that can be used routinely in cell culture laboratories to monitor cell line integrity.
The method involves the isoenzymatic separation of nine polymorphic enzymes, three of which can be used for cell line species
determinations and seven of which can be used for human cell line characterizations. Examples of how the system has been applied
to both inter- and intraspecies identifications are described. The routine application of this protocol would be a valuable
asset for laboratories concerned with establishing effective cell culture quality control.
This work was supported by Contract N01-CP-9-1003 from the National Cancer Institute, Bethesda, MD. 相似文献
12.
In a systematic effort aimed at identifying new steroidal cytotoxic agents with potent antiproliferative activity against cancer cells, we synthesized certain 16-[4-(NO2, CN, and i-Pr)substituted]benzylidene derivatives of androst-5-ene, 7-25, with pyrrolidino functionality in the 3beta-position of the steroid nucleus, i.e., 13-18 and 25. The selected compounds were examined for their cytotoxicity against a panel of three human cancer cell lines at the National Cancer Institute (NCI), Bethesda, USA. The results presented herein provide experimental evidence that compounds 7, 9, 10, 12, 16, and 19-21 induced apoptosis in human cancer cells. 相似文献
13.
Summary In this report we describe a new apparatus which has been developed for the automated selective dissociation of multicellular
spheroids into fractions of viable cells from different locations in the spheroid. This device is based on the exposure of
spheroids to a 0.25% solution of trypsin under carefully controlled conditions, such that the cells are released from the
outer spheroid surface in successive layers. Study of the spheroid size, number of cells per spheroid, and sections through
the spheroid with increasing exposure to trypsin demonstrate the effectiveness of this technique. The technique has been successfully
used on spheroids from five different cell lines over a wide range of spheroid diameters. We also present data detailing the
effect of varying the dissociation temperature, the mixing speed, the trypsin concentration, and the number of spheroids being
dissociated. The new apparatus has several advantages over previous selective dissociation methods and other techniques for
isolating cells from different regions in spheroids, including: a) precise control over dissociation conditions, improving
reproducibility; b) short time to recover cell fractions; c) ability to isolate large numbers of cells from many different
spheroid locations; d) use of common, inexpensive laboratory equipment; and e) easy adaptability to new cell lines or various
spheroid sizes. Applications of this method are demonstrated, including the measurement of nutrient consumption rates, regrowth
kinetics, and radiation survivals of cells from different spheroid regions.
This work was supported by grants CA-36535, CA-22585, and RR-02845 from the National Institutes of Health, Bethesda, MD, the
National Flow Cytometry Resource (NIH grant RR-01315), and by the Department of Energy, Washington, DC. 相似文献
14.
Development of two cloned epithelial cell lines from normal adult mouse and rat ventral prostates 总被引:1,自引:0,他引:1
Yoshinobu Kubota Eugene B. Gehly Karl H. Link Charles Heidelberger 《In vitro cellular & developmental biology. Plant》1981,17(11):965-978
Summary Two epithelial cell lines were established, one from adult C3H mouse and one from adult Fischer rat ventral prostate. These
cell lines were obtained from explant cultures, using Ham's F12 medium supplemented with HEPES, insulin, testosterone, hydrocortisone,
epidermal growth factor, and 7.5% fetal bovine serum. A low concentration of trypsin and EDTA in Ca++-and Mg++-free phosphate buffer was used for passaging the cells. The rat cell line was established following implantation of prostate
tissue in nude mice. These cell lines stained positively for acid phosphatase and were dependent upon epidermal growth factor
for growth. Morphological studies, including electron microscopy, revealed a highly characteristic epithelial morphology of
both cell lines. These cell lines have hypotetraploid chromosome numbers and are capable of metabolizing benzo(a)pyrene. We propose the application of these cells as models for the study of prostate carcinogenesis.
This work was supported in part by Grant CA-21, 746, and by the Electron Microscope Core Facility on Grant CA-14,089, from
the National Cancer Institute, National Institutes of Health, Bethesda, MD. 相似文献
15.
Alfonso Gonzalez Terry D. Oberley Janice L. Schultz Jennifer Ostrom Jonathan J. Li 《In vitro cellular & developmental biology. Animal》1993,29(7):562-573
Summary Primary diethylstilbestrol-induced kidney tumors from Syrian hamsters were grown in vitro and maintained in culture for 6
mo. Combined immunohistochemical studies using antibodies to intermediate filaments and ultrastructural studies of tumor cells
in culture exhibited characteristics similar to tumor cells in vivo. Furthermore, the cells manifested transformed properties
in culture; they grew both as multilayered colonies attached to the tissue culture substrate and as floating multicellular
colonies (spheroids). When cultured cells were injected into diethylstilbestrol-treated recipient hamsters, tumors developed
at the injection sites. In contrast, renal tubules or whole kidney cortex from control hamsters cultured in the same medium
underwent only short-term growth, with senescence developing after approximately 1 mo. However, cell cultures of kidney cortex
from animals treated in vivo for 5 mo. with diethylstilbestrol formed a cell line. This diethylstilbestrol-induced cell line
has been maintained in culture for 1.5 yr and has the following characteristics: a) it is anchorage-dependent, b) it is negative
in in vivo tumorigenicity tests, and c) cultured cells are histochemically and ultrastructurally similar to cultured tumor
cells. This culture system should prove to be of use in studying hormonal carcinogenesis in vitro.
This study was supported by the Medical Research Service, Department of Veterans Affairs, Washington, DC, and by grant CA-22008
from the National Cancer Institute, NIH, DHHS, Bethesda, MD. 相似文献
16.
J. R. Wagle J. J. Heindel A. Steinberger B. M. Sanborn 《In vitro cellular & developmental biology. Plant》1986,22(6):325-331
Summary Commonly used enzymic methods for the isolation of rat Seroli cells yield populations containing ∼15% germ cells. Although
the germ cells become eliminated after several media changes, they could interferen with the use of Sertoli cells for critical
studies during the first several days of culture. A brief treatment of Sertoli cell monolayer cultures with 20 mM Tris-HCl (pH 7.4) was found to eliminate most of the residual contaiminating germ cells. The duration of this treatment varied
from 1.0 to 10 min, depending on cell denisty in the culture, the degree of germ cell contamination, and the age of animals
used for Sertoli cell isolation. In a study of 95% pure 7-d Sertoli cell cultures, the hypotonic treatment did not alter the
DNA or RNA content per dish or the incorporation of [3H]uridine into total and poly A+RNA. Also, the hypotonic treatment did not alter specific Sertoli cell functions, i. e., secretion
of Sertoli cell factor (inhibin) and stimulation of cAMP levels by follicle stimuting hormone in 2-d cultures. Androgen receptor
concentration per dish was also not changed. Changes in several general metabolic parameters observed after hypotonic treatment
of 2-d cultures were attributed primarily to loss of contaiminating germ cells. Consequently, hypotonic treatment can be used
to eliminate contaminating germ cells from the Sertoli cell cultures without apparent detrimental effects on a number of Sertoli
cell biochemical parameters. This may be of considerable importance when the purity of Sertoli cells is critical for the interpretation
of experimental data.
This work was supported in part by grants HD-1-P50-08338, HD-17795 (BMS), and HD-18186 (JJH) from the National Institutes
of Health, Bethesda, MD. 相似文献
17.
Robert J. Klebe 《In vitro cellular & developmental biology. Plant》1984,20(2):127-132
Summary A rapid and technically simple method for cloning both adhesive and nonadhesive mammalian cells is described. The procedure
employs (a) honeycomb cloning plates and (b) nonlethal vital stains. Instead of placing cloning rings around colonies, cells
are initially seeded at clonal density directly into a plate containing an array of cloning rings (the honeycomb plate). Hence,
the time involved in placing cloning rings around colonies is eliminated. Second, clone-containing wells of the honeycomb
plate are easily identified by staining plates with the nonlethal vital stains, MTT or INT tetrazolium. Vital staining eliminates
the time involved in searching for clones. Last, clones are transferred with a cotton-tipped swab thereby eliminating the
time involved in trypsinization of cells. In this fashion, one can pick and transfer clones ofsubstrate adherent mammalian
cells at a rate of one clone/ 10 to 15 s. Thus, mammalian cells can be cloned as rapidly as cloning can be carried out in
microbial systems.
This study was supported, in part, by Grant CA 33074 from the National Cancer Institute, Bethesda, MD, Grant PCM-8218137 from
the National Science Foundation, Washington, D.C., and a grant from the National March of Dimes. 相似文献
18.
19.
Cultured human foreskin fibroblasts produce a factor that stimulates their growth with properties similar to basic fibroblast growth factor 总被引:2,自引:0,他引:2
Michael T. Story 《In vitro cellular & developmental biology. Plant》1989,25(5):402-408
Summary To determine if fibroblasts could be a source of fibroblast growth factor (FGF) in tissue, cells were initiated in culture
from newborn human foreskin. Cells were studied in Passages 2 to 8. Fibroblast cell lysates promoted radiolabeled thymidine
uptake by cultured quiescent fibroblasts. Seventy-nine percent of the growth-promoting activity of lysates was recovered from
heparin-Sepharose. The heparin-binding growth factor reacted on immunoblots with antiserum to human placenta-derived basic
FGF and competed with iodinated basic FGF for binding to antiserum to (1-24)bFGF synthetic peptide. To confirm that fibroblasts
were the source of the growth factor, cell lysates were prepared from cells incubated with radiolabeled methionine. Heparin
affinity purified material was immunoprecipitated with basic FGF antiserum and electrophoresed. Radiolabeled material was
detected on gel autoradiographs in the same molecular weight region as authentic iodinated basic FGF. The findings are consistant
with the notion that cultured fibroblasts express basic FGF. As these cells also respond to the mitogen, it is possible that
the regulation of their growth is under autocrine control. Fibroblasts may be an important source of the growth factor in
tissue.
Supported in part by grant DK 31063 from the National Institutes of Health, Bethesda, MD. 相似文献
20.
Summary Two commercially available serum replacements developed for use in the culture of hybridoma and other mammalian cells were
tested for their suitability as replacements for fetal bovine serum in insect cell culture medium. CPSR-1 and CPSR-3 both
supported growth of the insect cell line IPLB-SF-21AE. CPSR-3 supported adequate growth, but cells in medium supplemented
with CPSR-1 grew much slower and achieved only about half the final cell density of either FBS or CPSR-3 supplemented medium.
This work was supported in part by grant 187159 from the Juvenile Diabetes Foundation and BRSG RR05876 from the National Institutes
of Health, Bethesda, MD. 相似文献