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1.
Lawrence E. DeBault Larry E. Kahn Stephen P. Frommes Pasquale A. Cancilla 《In vitro cellular & developmental biology. Plant》1979,15(7):473-487
Summary Microvessels isolated from mouse forebrain were used as the source material for the derivation of cerebral vascular endothelium
and smooth-muscle cells in culture. The microvessels were isolated by a mechanical dispersion and filtration technique, and
were maintained in vitro as organoid cultures. A microvessel classification system was developed and proved to be useful as
a tool in monitoring culture progress and in predicting the type(s) of microvessel(s) that would give rise to migrating and/or
proliferating cells. The isolated cerebral microvessels were heterogeneous in diameter, size of individual vascular isolate,
and proliferative potential. The isolated microvessels ranged in diameter from 4 μm to 25 μm and in size from a single microvascular
segment to a large multibranched plexus with mural cells. The initial viability, determined by erythrosin B exclusion, was
approximately 50% on a per cell basis. All microvessel classes had proliferative potential although the rate and extent of
proliferation were both microvessel class- and density-dependent. The smaller microvessels gave rise to endothelial cells,
whereas the large microvessels gave rise to endothelial and smooth-muscle cells. The viability and progress of a microvessel
toward derived cell proliferation seemed to be directly proportional to the number of mural cells present.
This work was supported in part by an Arteriosclerosis Specialized center of Research grant from the National Heart, Lung
and Blood Institute, National Institutes of Health (HL-14230) and Grant 584-127703 from the Veterans Administration. 相似文献
2.
Boyd R. Switzer George K. Summer 《In vitro cellular & developmental biology. Plant》1973,9(3):160-166
Summary In human diploid skin fibroblasts in culture we have shown that nonhydroxylated collagen precursors remain in the cell when
proline hydroxylation is inhibited by α, α′-dipyridyl, a chelator of ferrous ions. The inhibition of proline hydroxylation
is reversed by addition of fresh medium containing 50 μg per ml of sodium ascorbate, whereupon nonhydroxylated collagen precursors
are hydroxylated within the cell and extruded into the medium. Extrusion of collagen already formed within the cell is not
appreciably affected by α, α′-dipyridyl inhibition. Under normal conditions collagen is released from the monolayer into the
medium within 3 hr of a pulse ofL[14C]proline. In the presence of α, α′-dipyridyl, about 35% of theL[14C]proline incorporated into protein is released into the medium within 8 hr as a proline-rich, hydroxyproline-deficient protein;
at the same time, approximately 15% of the protein-boundl-[14C]proline remains in the cell for as long as 12 hr. When proline hydroxylation is restored after 2 and 12 hr of α, α′-dipyridyl
inhibition, approximately the same amount of hydroxyproline is formed after each time interval in the monolayer. Therefore,
nonhydroxylated collagen precursors retained in the cell are not appreciably degraded during at least 12 hr of inhibition
by α, α′-dipyridyl and are extruded into the medium only upon restoration of hydroxylation.
This work was supported in part by a grant from the Easter Seal Research Foundation, and by Project 236, Health Services and
Mental Health Administration, Department of Health, Education and Welfare, Grant HD-03110 from the National Institute of Child
Health and Human Development, an American Cancer Society Institutional Grant (1N 15-J), a General Research Support Award (5-S01-FR-05406)
from the National Institutes of Health, a University Research Council Grant, a National Science Foundation Equipment Grant
(GB-4577), and a Research Career Development Award (5-K3-AM-5058) from the National Institute of Arthritis and Metabolic Disease
(G.K.S.). 相似文献
3.
Philip Coffino Henry R. Bourne Paul A. Insel Kenneth L. Melmon Gary Johnson Josefina Vigne 《In vitro cellular & developmental biology. Plant》1978,14(1):140-145
Summary S49 mouse lymphoma cell mutants, each with a specific defect in its ability to generate or respond to cyclic AMP, have been
isolated. Analysis of the properties of these cells has begun to provide information on complex and significant biologic problems
related to the cyclic AMP system.
Presented in the Opening Symposium on Nutritional Factors and Differentiation at the 28th Annual Meeting of the Tissue Culture
Association, New Orleans, Louisiana, June 6–9, 1977.
The work was supported in part by National Science Foundation Grant BMS 75-06764 and National Institutes of Health Grants
GM 16496 and GM 00001. P.C. is the recipient of National Institutes of Health Research Career Development Award GM 00308.
P. A. I. is an Established Investigator of the American Heart Association. 相似文献
4.
Lawrence E. DeBault Eduardo Henriquez Michael N. Hart Pasquale A. Cancilla 《In vitro cellular & developmental biology. Plant》1981,17(6):480-494
Summary An endothelial cell line has been established from a primary culture of cerebral microvessels isolated from Swiss-Webster
mice. The microvessels were isolated by a mechanical dispersion and filtration technique. The cells that emerged from these
microvessels, maintained in organoid cultures, proliferated and formed plaques of a single or mixed cell type. The endothelial
cell line, designated ME-2, was isolated from one such morphologically homogeneous cell plaque, using both cloning ring techniques
and C6 glioma-conditioned medium.
An endothelial specific antiserum was made in rabbits and was used immunocytochemically to confirm the cell type of origin
of the ME-2 cell line. Not only did the cell type specific antiserum react exclusively with endothelial cells in vivo, but
in the brain the antiserum localized preferentially to the luminal membrane of the endothelium.
The ME-2 endothelial cells have retained several of their unique properties such as cytomorphology, growth characteristics,
and cell type specific surface antigens throughout the life of the line (in one case 40 passages before senescence).
This work was supported in part by an Arteriosclerosis Specialized Center of Research grant from the National Heart, Lung
and Blood Institute, National Institutes of Health, Grant HL-14230, and Grant 584-127703 from the Veterans Adminsitration.
This paper is dedicated to the memory of Steve Frommes, Electron Microscopist and Photographer. 相似文献
5.
Dr. Juliet Morgan Louis Cohen 《In vitro cellular & developmental biology. Plant》1974,10(3-4):188-195
Summary This report describes in detail a method of enzymatic separation of adult mammalian muscle using papain. The procedure has
proved valuable in the preparation of suspensions of muscle cell pieces from normal human skeletal muscle obtained from patients
of all ages, from 3 months to 79 years. Muscle cultures have been successfully growth from biopsy material from boys with
Duchenne’s muscular dystrophy and from their mothers. The procedure was initially established with adult canine skeletal muscle
and has also been used for monkey muscle.
Small pieces of skeletal muscle are chopped in a solution of 0.05% papain and 0.01% cysteine hydrochloride in Ca2+-and Mg2+-free balanced salt solution and transferred in the papain solution to a flask, in which they are incubated at 37°C for 10
min with occasional agitation. The resulting cell suspension is collected and the remaining pieces are treated with further
portions of fresh papain until only connective tissue remains. The cell pellets obtained by centrifugation are resuspended
in Eagle’s minimum essential medium (supplemented with 20% fetal calf serum) and transferred to culture chambers. The muscle
can be observed at all times, during the separation procedure and subsequently in culture. The events occurring during skeletal
muscle regeneration can be followed. Using the same papain preparation, myoblasts and myotubes may be subcultured and collected
for indefinite frozen storage in dimethylsulfoxide.
This work was supported by a grant-in-aid from the American Heart Association with funds contributed in part by the Chicago
Heart Association, the Pharmaceutical Manufactures Association, and National Institutes of Health Research Grant NS 10385
from the Institute of Neurological Diseases and Stroke. 相似文献
6.
Gordon Parry Debble Farson Betsey Cullen Mina J. Bissell 《In vitro cellular & developmental biology. Plant》1988,24(12):1217-1222
Summary Primary cultures of mouse mammary epithelial cells synthesize significant quantities of chondroitin and heparan sulfate proteoglycans
(16). Long term treatment of such cultures with p-nitrophenyl-β-D-xylopyranoside leads to a 10–20 fold increase in the synthesis
and secretion of free chondroitin sulfate glycosaminoglycan (GAG) chains and assembly of a cell-associated matrix that is
relatively enriched in heparan sulfate proteoglycan.
This modulation of cell-synthesized proteoglycans leads to significant changes in cell morphology and cellular differentiation.
Notably cells cultured on plastic culture dishes change from being flattened to cuboidal. The synthesis of the milk proteins
α1, α2, and β-casein is also increases as is the formation of fat droplets and fat droplet membrane components. Promotion
of differentiation increases with increasing xyloside concentration in the range 0–1.5 mM, but there may be a block in secretion
at higher xyloside concentrations.
While the detailed mechanisms remain to be elucidated, we conclude that the composition of proteoglycans incorporated into
the matrix (and possibly the glycosaminoglycans secreted into the medium), may play a significant role in maintaining the
phenotypic characteristics of terminally differentiated mammary epithelial cells.
This research was supported by the Office of Health and Environmental Research, Office of Energy Research, U.S. Dept. of Energy
under contract No. DEAC-03-76SF00098 and by National Institutes of Health Grant CA44398-01 (G. Parry)
Editor's Statement Exogenous elements of extracellular matrix affect expression of cultured mammary cell function. This work
reports manipulation of cell-derived endogenous matrix elements and shows correlative changes in cell functions. 相似文献
7.
Joseph L. Melnick Janet S. Butel Satvir S. Tevethia Nilambar Biswal Matilda Benyesh-Melnick 《In vitro cellular & developmental biology. Plant》1971,6(5):349-354
Summary This paper describes some current work pertaining to transformation of cells by oncogenic viruses.
Part I includes: (1) the effect of a deoxyribonucleic acid (DNA) tumor virus (SV40) on the antigenic characteristics of transformed
cells; (2) in vitro and in vivo methods of detecting virus-specific surface antigens; (3) the role that the host cell may
play in the expression of virus-coded antigens; and (4) the presence of virus-induced antigens as a possible mechanism of
the apparent nononcogenicity of certain virus variants.
Part II discusses (1) the physicochemical properties of the nucleic acid of a ribonucleic acid (RNA) tumor virus-the Moloney
sarcoma-leukemia virus (MSV-MLV) complex —(2) a preliminary analysis of viral RNA replication in cells transformed by MSV-MLV,
and (3) application to human tumors.
Supported in part by Research Grant CA 04600 and by Research Contract PH 43-68-678 within the Special Virus-Cancer Program,
National Cancer Institute, National Institutes of Health.
Recipient of Research Career Development Award 5-K3-CA 38,614 from the National Cancer Institute, National Institutes of Health 相似文献
8.
Isolation and culture of cells derived from human cerebral microvessels 总被引:10,自引:0,他引:10
Harry V. Vinters Susan Reave Penny Costello John P. Girvin Steven A. Moore 《Cell and tissue research》1987,249(3):657-667
Summary Microvessels were isolated from non-neoplastic human cerebral cortical fragments resected for treatment of intractable seizure disorder. The microvessels were incubated in modified Lewis medium with 20 or 30% fetal bovine serum. Within 1–2 weeks, two cell populations emerged from the isolates. One type of cells had polygonal morphology, showed density-dependent contact inhibition at confluence in vitro, showed lectin-binding characteristics of endothelium (but only moderate positivity for factor VIII antigen), demonstrated induction of -glutamyl trans-peptidase when exposed to astrocyte-conditioned media, and responded to insulin by a pronounced increase in DNA synthesis. The other variety of cells grew in vitro more slowly in irregular strands separated by clear zones, showed ultrastructural features of smooth muscle, and isoelectric focusing of cell proteins revealed the presence of smooth-musclespecific -isoactin. Both types of cells could be serially subcultured. The ability to isolate and grow the two cell types, tentatively identified as human cerebral microvascular endothelium and smooth muscle, may facilitate studies of human blood-brain barrier function as well as the pathogenesis of cerebral microangiopathies unique to the human brain.Funded by Canadian Heart Foundation, Heart and Stroke Foundation of Ontario and UCLA Biomedical Research Support Grant 相似文献
9.
C. D. Thron 《Bulletin of mathematical biology》1982,44(5):609-635
This paper deals with the unimodality of the ‘impulse response’ in compartmental systems, where the ‘impulse response’ in
any given compartment is the time course of the amount of diffusing substance in that compartment after an initial instantaneous
injection of the substance into that or some other compartment. It is shown that in certain compartmental structures, with
injection in certain compartments, the impulse response is always unimodal or monotonic in all compartments, regardless of
the numerical values of the various transfer rate coefficients. Structures with this property are here named ‘UM structures’,
and they include the familiar mammillary and catenary structures. In this paper, the set of all UM structures is described.
Structures which are not UM (NUM structures) are identified by showing that, by removal of certain connections and compartments
according to certain rules, they can be reduced to small structures which can be shown to be NUM by numerical computation.
Computations on two systems with bimodal impulse responses show that with constant infusions of a fixed amount of substance
the peak level may increase paradoxically with decrease in the infusion rate over a certain range. This effect is extremely
small, however.
Supported in part by U.S. Public Health Service Research Grant GM 21269 from the National Institute of General Medical Sciences,
and in part by Biomedical Research Support Grant S07 RR 05392 from the National Institutes of Health. 相似文献
10.
Steve Hamner Walis Jones Jean R. Starkey Howard L. Hosick 《In vitro cellular & developmental biology. Plant》1989,25(12):1107-1113
Summary Three related mouse mammary cell lines were cultured in collagen gels and assayed for growth factor responsiveness and interaction
via soluble factors. The CL-S1 cell line is nontumorigenic and grows poorly in collagen gel culture. The +SA and −SA cell
lines exhibit different degrees of malignant behavior in vivo and have different growth properties in vitro. In collagen gel
culture, +SA growth was stimulated by serum but not by epidermal growth factor (EGF), whereas both serum and EGF were required
for optimal growth of −SA cells of early passage number as well as CL-S1 cells. −SA cells of later passage repeatedly exhibited
a change so as to no longer require serum while retaining EGF responsiveness. [125I]EGF binding analyses indicated that CL-S1 cells bound EGF with less affinity than did −SA cells whereas +SA cells bound
almost to ligand. When cell lines were maintained in separate collagen gels but shared the same culture medium, growth of
+SA or −SA cells was slightly enhanced in the presence of CL-S1 cells and −SA cell growth was enhanced by the presence of
+SA cells. Using the normal rat kidney fibroblast line NRK (clone 49F) as an indicator, serum-containing conditioned media
from each cell line and from each pair of cell lines cultured in collagen gels were tested for transforming growth factor
(TGF) activity. Both the −SA and CL-S1 lines tested positive for TGF-α production and possibly released a TGF-β activity.
These results suggest mechanisms by which cell populations in and around tumors can modify one another’s growth characteristics.
The work was supported by a grant from the American Institute for Cancer Research, by American Cancer Society Institutional
grant IN-119, by funds from the Poncin Trust (Seattle-First National Bank), and by grants CA-39611 and CA46885 from the National
Institutes of Health, Bethesda, MD. 相似文献
11.
Isolation,characterization, and long-term culture of fetal bovine tracheal epithelial cells 总被引:5,自引:0,他引:5
Brenda L. Schumann Terence E. Cody Marian L. Miller George D. Leikauf 《In vitro cellular & developmental biology. Plant》1988,24(3):211-216
Summary Epithelial cells were isolated from fetal bovine trachea by exposing and stripping the mucosal epithelium from the adjacent
connective tissue. The tissue was minced and enzymically dissociated in Ca-Mg-free medium containing dispase and dithiothreitol.
The stripping procedure and selective trypsinization produced epithelial cell cultures free of fibroblasts. Seeded on plastic,
the plating efficiency was 21.5% with a doubling time of 24 h. Dome formation, evidence of occluding junctions and active
ion transport characteristic of epithelial cells, was common. Growth of the cells on glass, collagen, and Engelbreth-Holm-Swarm
(EHS) substrate demonstrated a striking difference in morphology. Cells grown on EHS presented a more distinctly three-dimensional
growth pattern and many more microvilli when compared to cells grown on glass or collagen. The cells retained their epithelioid
characteristics through more than 30 passages as shown by the presence of distinct apical and basolateral membranes, tight
junctions, and positive keratin staining.
This study was supported in part by grants BRSG S07 RR05408-25, Biomedical Research Support Grant Program, Division of Research
Resources, by ES 00159, Center Grant, National Institutes of Environmental Health Sciences, by R23-HL37621, New Investigator
Award, National Heart, Lung and Blood Institutes, National Institutes of Health, and by the Health Effects Institute, an organization
jointly funded by the U.S. Environmental Protection Agency (Assistance Agreement X-8120059) and automotive manufactures. The
contents do not necessarily reflect the views of policies of HEI, EPA, or automotive manufacturers. 相似文献
12.
Arthur E. Greene Jesse Charney Warren W. Nichols Lewis L. Coriell 《In vitro cellular & developmental biology. Plant》1972,7(5):313-322
Summary Three mosquito cell cultures designated as Suitor's clone ofAedes aegypti, Culiseta inornata, andAedes vexans were shown to be moth by immunological, karyological, and isozyme analyses. The cells reacted with rabbit antimoth serum
but not rabbit antimosquito serum. Chromosome analyses indicated Lepidopteran rather than Dipteran morphology, and three isozyme
systems were confirmative. Any one of these assays would be sufficient to indicate that contamination had occurred and could
be used as a periodic check for identity of cell cultures. Morphology and growth characteristics are also valid criteria to
distinguish between these particular orders of insect cells.
These studies were supported by Grant CA-04953-12 from the National Cancer Institute; General Research Support Grant FR-5582
from the National Institutes of Health; and Grant-in-Aid Contract M-43 from the State of New Jersey.
Recipient of Research Career Award 5-K3-16,749 from the National Institutes of Health. 相似文献
13.
To examine whether differences in chondrocytes from skeletally immature versus adult individuals are important in cartilage
healing, repair, or tissue engineering, superficial zone chondrocytes (SZC, from within 100 μm of the articular surface) and
deep zone chondrocytes (DZC, from 30%–45% of the deepest un-mineralized part of articular cartilage) were harvested from immature
(1–4 months) and young adult (18–36 months) steers and compared. Cell size, matrix gene expression and protein levels, integrin
levels, and chemotactic ability were measured in cells maintained in micromass culture for up to 7 days. Regardless of age,
SZC were smaller, had a lower type II to type I collagen gene expression ratio, and higher gene expression of SZ proteins
than their DZC counterparts. Regardless of zone, chondrocytes from immature steers had higher levels of Sox 9 and type II
collagen gene expression. Over 7 days in culture, the SZC of immature steers had the highest rate of proliferation. Phenotypically,
the SZC of immature and adult steers were more stable than their respective DZC. Cell surface α5 and α2 integrin subunit levels
were higher in the SZC of immature than of adult steers, whereas β1 integrin subunit levels were similar. Both immature and
adult SZC were capable of chemotaxis in response to fetal bovine serum or basic fibroblast growth factor. Our data indicate
that articular chondrocytes vary in the different zones of cartilage and with the age of the donor. These differences may
be important for cartilage growth, tissue engineering, and/or repair.
This work was supported in part by the National Chapter of the Arthritis Foundation, the Ira DeCamp Fellowship for Musculoskeletal
Research of the Hospital for Special Surgery, the Institute for Sports Medicine Research in New York, and the National Institute
of Health grant AR045748 and was conducted in a facility constructed with support from the Research Facilities Improvement
Program (grant no. C06-RR12538-01) of the National Center for Research Resources, National Institutes of Health. 相似文献
14.
Cultured circular smooth muscle from the rabbit colon 总被引:1,自引:0,他引:1
H. W. Kao S. E. Finn A. M. Gown J. Lechago N. Lachant W. J. Snape Jr. 《In vitro cellular & developmental biology. Plant》1988,24(8):787-794
Summary Although cultured vascular smooth muscle cells have been extensively characterized and investigated, there are very few studies
of cultured intestinal smooth muscle cells. The aim of this study was to culture colonic smooth muscle (CSM) cells from the
rabbit colon. Freshly isolated CSM cells from the circular muscle layer of the distal colon were prepared by collagenase digestion.
In primary culture, CSM cells attached to the culture vessels by 48 to 72 h, proliferated by 3 to 7 d, and reached confluency
by 14 to 17 d with a “hill-and-valley” pattern. Spontaneous contractions were not observed at any time at 21° or 37° C. Confluent
primary cultures were greater than 95% CSM cells, as identified by intensely positive immunofluorescent staining to smooth
muscle actin-specific CGA7 and muscle-specific HHF-35 monoclonal antibodies. Transmission electron microscopy of freshly isolated
and proliferating CSM cells revealed ultrastructural features consistent with smooth muscle cells. We successfully cultured
CSM cells of the rabbit from freshly isolated cells and validated these CSM cells by electron microscopy and immunocytochemical
staining. These highly pure primary cultures may be used to investigate numerous aspects of CSM cell metabolism and physiology.
These studies were supported by the National Institutes of Health grant to the Inflammatory Bowel Disease Center (Bethesda,
MD) P30-AM-32200 and R01-DK-31147. Dr. Kao is the recipient of a Research Career Development Award from the National Foundation
for Ileitis and Colitis, Inc. A preliminary report of this work was presented at the American Motility Society Meeting, Houston,
TX, in October 1986, and appeared in abstract form inGastroenterology 91: 1057; 1986. 相似文献
15.
Robert H. Hysmith T. Kenneth Welch Paul J. Boor 《In vitro cellular & developmental biology. Plant》1987,23(2):129-133
Summary This study measures the inhibition of [3H]uridine uptake by smooth muscle cells to determine sublethal toxic injury by the cardiovascular toxins, allylamine, isoproterenol,
and β-aminopropionitrile. The exposure period or the concentration of toxin which inhibited 30% of [3H]uridine uptake by smooth muscle cells could be utilized as an endpoint for ranking toxicity. Cytotoxicity of the three toxins
to smooth muscle cells were ranked as: allylamine > isoproterenol > β-aminopropionitrile. Recovery of cells utilizing [3H]uridine uptake inhibition as a method for assessing comparative cytotoxicity and for screening of agents potentially injurious
to vascular cells.
This work was supported by grant HL 26189 from the National Institutes of Health, Bethesda, MD and Research Career Development
Award HL 00929 (P. J. B.). 相似文献
16.
A method for the continuous cultivation of mammary epithelium 总被引:1,自引:0,他引:1
Etienne Y. Lasfargues Dan H. Moore 《In vitro cellular & developmental biology. Plant》1971,7(1):21-25
Summary Established cell lines of mammary epithelium have been obtained from mice, rats, and hamsters. Maintenance and replication
of the epithelium in serial subcultures were dependent on their periodic treatment with collagenase. Because collagenase is
not cytotoxic and has maximum efficiency at neutral pH in isotonic saline solutions containing calcium and magnesium, this
enzyme can be introduced directly into the culture medium; cells have been maintained for 3 days in such a medium with serous
enrichment at no detriment to them. Up to 10% concentrations of serum have not interfered with enzymatic activity.
Supported by United States Public Health Service Grant CA-08515 and CA-08740 from the National Cancer Institute. General Research
Support Grant FR-5582 from the National Institutes of Health, and Grant-in-Aid Contract M-43 from the State of New Jersey. 相似文献
17.
Arthur E. Greene Lorraine Toji Warren W. Nichols Lewis L. Coriell 《In vitro cellular & developmental biology. Plant》1973,9(3):156-159
Summary Forty-six cell cultures established from amniocentesis fluids were preserved in liquid nitrogen and later recovered from the
frozen state with little loss of viability as compared to prefreeze viability. Five to 10% glycerol was found to be optimal
for preservation in liquid nitrogen, and as few as 5×105 viable cells per frozen ampule could initiate cell growth. Storage in liquid nitrogen did not affect the genetic stability
of glucose-6-phosphate dehydrogenase, lactate dehydrogenase, malic dehydrogenase, leucine aminopeptidase, acid phosphatase,
or 6-phosphogluconic acid dehydrogenase isozymes of the amnion cultures.
These studies were supported by Contract NIH-NIGMS-72-2070, Grant CA-04953-13 from the National Cancer Institute; General
Research Support Grant FR-5582 from the National Institutes of Health; and Grant-in-Aid Contract M-43 from the State of New
Jersey.
Recipient of Research Career Award 5-K3,16, 749 from the National Institutes of Health. 相似文献
18.
Mary E. Riser Bonnie C. Huff Daniel Medina 《In vitro cellular & developmental biology. Plant》1983,19(9):730-734
Summary Normal mouse mammary epithelial cells in primary culture can be passaged as viable single cells using 0.5 to 1.0 mg/ml pepsin
in Hanks’ salt solution. After 5 min the pepsin treatment preferentially removes fibroblasts, leaving a monolayer of purified
epithelial cells that can be removed by pipetting and transferred to new culture vessels or injected into animals.
This study was supported by Contract CB-43907 from the National Institutes of Health. 相似文献
19.
Fibroblast spreading induced by connective tissue stretch involves intracellular redistribution of α- and β-actin 总被引:3,自引:3,他引:0
Langevin HM Storch KN Cipolla MJ White SL Buttolph TR Taatjes DJ 《Histochemistry and cell biology》2006,125(5):487-495
Mechanical stretching of connective tissue occurs with normal movement and postural changes, as well as treatments including physical therapy, massage and acupuncture. Connective tissue fibroblasts were recently shown to respond actively to short-term mechanical stretch (minutes to hours) with reversible cytoskeletal remodeling, characterized by extensive cell spreading and lamellipodia formation. In this study, we have examined the effect of tissue stretch on the distribution of α- and β-actin in subcutaneous tissue fibroblasts ex vivo. Normal fibroblasts uniformly exhibited α-smooth muscle actin (α-SMA) immunoreactivity. Unlike cultured fibroblasts and smooth muscle cells, α-SMA in these fibroblasts was not in F-actin form (indicated by lack of phalloidin co-localization) nor was it organized into distinct stress fibers. The lack of stress fibers and fibronexus was confirmed by electron microscopy, indicating that these cells were not myofibroblasts. In unstretched tissue, the pattern of α-actin was diffuse and granular. With tissue stretch (30 min), α-actin formed a star-shaped pattern centered on the nucleus, while β-actin extended throughout the cytoplasm including lamellipodia and cell cortex. This dual response pattern of α- and β-actin may be an important component of cellular mechanotransduction mechanisms relevant to physiologic and therapeutic mechanical forces applied to connective tissue. 相似文献
20.
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