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1.
Hamster early two-cell embryos developed to the expanded blastocyst stage within the isolated mouse ampulla maintained in organ culture system. Mouse ampullae isolated at different times after treating the mice with human chorionic gonadotropin (hCG) (0–72 h) or pregnant mare's serum gonadotropin (PMSG) (30–32 h) were flushed with culture medium, and hamster early two-cell embryos were introduced into these ampullae. Mouse ampullae isolated at 14–32 h after hCG injection were more favorable for the development of the embryos than those isolated at 70–72 h. When mouse ampullae were isolated 30–32 h after hCG or PMSG treatment, 39% of the cultured eggs developed, some of them to the expanded blastocyst stage after additional culture for 65–70 h. These results indicate that unknown oviductal factors stimulate the development of hamster early two-cell embryos, and these factors are under the control of hCG or PMSG. In addition, these factors are common to the mouse and hamster. 相似文献
2.
A. A. J. J. L. Rutten B. G. A. G. G. Béquet-Passelecq H. B. W. M. Koëter 《In vitro cellular & developmental biology. Plant》1990,26(4):353-360
Summary A new method was developed for rabbit skin organ culture. In a two-compartment model, skin discs were cultured on a Millicell-HA
insert unit with a microporous membrane which allows transport of culture medium via the dermis into the epidermis, whereas
the epidermal side remains free of direct contact with culture medium. In this relatively simple two-compartment organ culture
model, rabbit skin could be cultured for 7 d in RPMI 1640 medium supplemented with fetal bovine serum, or for 2 d in RPMI
1640 medium supplemented with cofactors. The histomorphology and ultrastructure of 7-d cultured rabbit skin discs was essentially
similar to that of freshly isolated rabbit skin. Keratinocytes in the stratum basale continued to divide during organ culture.
The terminal differentiation of the epidermis continued in vitro as was found by the presence of keratohyalin granules, the
intact stratum corneum, and keratin expression. Furthermore, glucose consumption continued until culture Day 7, but thereafter
it declined rapidly. Concomitantly, degenerative changes were found. At the end of the 7-d culture period the distance between
single dermal collagen fibrils had increased as compared to noncultured skin. This model of skin organ cultures can be used
to study biological processes, dermal toxicity, and penetration and metabolism of xenobiotics in intact skin. Furthermore,
within certain limits, processes responsible for repair and regeneration of damaged skin can also be studied in this model
because the rabbit skin can be cultured for 7 d.
The present study was financially supported by grants of Duphar B. V. (Weesp, Netherlands), the European Community, and the
Dutch animal welfare organizations Samenwerkingsverband van de Nederlandse Vereniging tot Bescherming van Dieren en de Nederlandse
Bond tot Bestrijding van de Vivisectie, Anti-Vivisectie Stichting en Stichting Schoonheid Zonder Wreedheid. 相似文献
3.
Retha A. Newbold Donald B. Carter Stephen E. Harris John A. McLachlan 《In vitro cellular & developmental biology. Plant》1981,17(1):51-54
Summary This report describes a serum-free system for studying the fetal mouse genital tract in organ culture. Using the techniques
of isoelectric focusing and gel electrophoresis of polypeptides synthesized by the organ explant in culture, the biochemical
integrity of as little as 1 to 2 mg of this tissue was determined during the period of culture. Thus, differentiation of the
fetal genital tissue in organ culture can be assessed by both morphological and biochemical criteria. 相似文献
4.
Summary A modified continuous-flow culture system (CFCS) was developed to maintain large explants of periodontium from adult mouse
in organ culture. The culture medium was stored in a reservoir outside of the incubator, pumped via polyvinyl tubing into
small glass culture chambers that were placed in the oxygenator and then collected in a waste flask. Medium was analyzed for
pO2, pCO2 and pH during the culture period. Three-molar and singlemolar explants of periodontium were maintained for 48 hr in the CFCS
at two different pO2 ranges: 100 to 120 mm Hg and 400 to 420 mm Hg. [3H]Proline was added 24 hr prior to sacrifice. Light-microscope morphological and radioautographic observations suggested that
cell viability and incorporation of [3H]proline, probably into newly synthesized protein, increased with an increase in pO2 and was related to a pO2 gradient extending from the periphery to the center of the explants. 相似文献
5.
G. Calaf I. H. Russo L. D. Roi J. Russo 《In vitro cellular & developmental biology. Plant》1986,22(3):135-140
Summary Several studies have shown the importance of different hormones in the regulation of mammary tsssue growth. The use of organ
culture techniques has shown tremendous value for the knowledge of cell proliferation in human breast tissue. Therefore, the
purpose of these studies was to analyze the length of the cell cycle, DNA-labeling index, mitotic index, and growth fraction
under the effects of insulin, hydrocortisone, and 17-β estradiol in 5-d organ culture. Normal tissues obtained from patients
who underwent breast surgery for benign lesions were individually cultured at 37°C (95% air:5% CO2 in Medium 199). Autoradiographic studies indicated that the hormones shortened the length of cell cycle of normal breast
tissue in 5-d organ cultures. From the growth fraction studies we concluded that the hormones may have stimulated the cells
to reenter the cell cycle from G0 because these values were increased by the hormones used. Estrogen can alter the S phase duration with a consequent increase
in the rate of DNA synthesis which may explain the high DNA-labeling index observed in the present studies.
Supported by Public Health Service grant CA38921 from the National Cancer Institute, Bethesda, MD, and by an Institutional
grant from the United Foundation of Greater Detroit. 相似文献
6.
Summary The effect of variations in culture conditions on survival of fragments of mouse and rat descending colon in organ culture
was studied by morphological and functional criteria. A combination of conditions demonstrated to be beneficial permitted
maintenance for at least 35 days. These included: a gaseous environment of 95% O2:5% CO2, an attachment matrix consisting of a Metricel GA-4 membrane (pore size, 0.8 μ), intermittent exposure to the gas and fluid
phases by rocking in 5 ml medium and supplementation of the medium with 1.0 μM dexamethasone and 10% FBS. During this time,
the crypt structure of the mucosal epithelium was well preserved, and DNA synthesis in the crypts and mucin production in
the crypts and superficial epithelium continued. In addition, the synthetic hormone, pentagastrin, stimulated DNA synthesis
in the mucosal epithelium of mouse colon fragment in short-term organ culture.
This work was supported by Environmental Protection Agency Grant R803998-01-1 and National Cancer Institute Contract N01-CP-75952. 相似文献
7.
Herman Yeger Diane Forget Jennifer Alami Bryan R. G. Williams 《In vitro cellular & developmental biology. Animal》1996,32(8):496-504
Summary The temporal and spatial expression patterns of the Wilms tumor gene, WT1, were studied during the organogenesis of the mouse
kidneyin vitro. In situ hybridization and immunocytochemistry localized cellular expression of WT1 in whole kidney organ cultures to the induced
metanephric mesenchyme and developing podocytes. Organ cultures were further characterized immunocytochemically with antibodies
that specifically labeled the different tubular epithelial components and supporting mesenchyme of the developing nephrons.
In organ cultures, the WT1 expression pattern could be visualized in induced metanephric mesenchyme and entire cell cohorts
of differentiating podocytes. Expression of WT1 and cell specific markers were retained in short-term monolayer cultures of
dissociated kidneys. The development of the metanephric kidneyin vitro involves a highly restricted temporal and spatial cellular expression pattern of WT1 which closely follows that observed
in tissue sections from gestational kidney isolated during organogenesis in the mouse. 相似文献
8.
Masayoshi Kumegawa Taishin Takuma Fumiko Murayama 《In vitro cellular & developmental biology. Plant》1976,12(10):718-728
Summary A new technique for organ culture which uses plastic culture chambers and the advantages of the cellophane-sheet technique
is described with the results of a study of cultivations of fetal mouse liver. Two chambers, each containing cells, were placed
in gas permeable roller tubes and rotated at 0.1 rpm in a CO2-air gassed incubator. The fetal mouse liver cells developed electron microscopic features similar to those of the in vivo
adult liver by 9 days of cultivation. The albumin content and tyrosine aminotransferase (TAT) activity were detected in the
cultivated liver. TAT activity was further induced by prednisolone. These results indicate the potential of this culture method
for the study of physiological and pathological processes.
This work was supported in part by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science and Culture,
Japan and Science Technology Agency, Japan. 相似文献
9.
Melania Maglio Matilde Tschon Laura Sicuro Roberta Lolli Milena Fini 《Journal of cellular physiology》2019,234(5):5420-5435
The increasing demand for reliable preclinical models and to reduce, refine and, if possible, replace animal studies have brought forth the development of complex tissue cultures in different research areas, including the musculoskeletal field. In this paper, we review the literature within last 10 years on the state of progress for in vitro models of osteochondral tissue cultures, taking into account the clinical relevance of the management and treatment of osteochondral lesions. According to the selected research criteria, 35 works, 27 of which with animal tissues and 8 with human tissues, resulted to be relevant for the purposes of this review. Data analyzed revealed a great heterogeneity among the proposed tissue culture models. The anatomical harvesting sites resulted to be mainly the knee stifle joint, both for animal (prevalently bovines) and human tissues derived from joint replacement surgery, and significant heterogeneity among culture conditions and media were found. To date, very few papers have focused on the set up of a reproducible in vitro model, applicable to a variety of studies, thus suggesting a relevant gap to fill in the development of advanced three-dimensional osteochondral culture models. 相似文献
10.
Judy M. Strum Elizabeth A. Hillman 《In vitro cellular & developmental biology. Plant》1981,17(1):33-43
Summary Normal breast tissue from a 17-year-old girl was grown in organ culture for 3 weeks. A comparison was made between the effects
on the epithelium of a defined culture medium containing various combinations of hormones and a serum-supplemented medium
that has been used to successfully maintain other human tissues for 4 months routinely, and in some cases for up to 1 year.
After culture for 3 weeks the explants were exposed to [3H]thymidine and autoradiographs were prepared and evaluated in order to determine labeling indexes. The only serum-free defined
medium that permitted any significant survival or labeling of the cells contained insulin + hydroxycortisone + prolactin.
However, serum-supplemented medium alone gave an even higher labeling index, and this was elevated more in media containing
either progesterone or other combinations of hormones. Our study indicates that normal human breast (removed at the early
postovulatory stage of the menstrual cycle) can be maintained in a differentiated state for 12 days in serum-supplemented
media. By 2 weeks the cells had begun to migrate onto the surface of the explant. They then began to accumulate tonofilaments
so that after 3 weeks in culture nearly all of the cells contained tonofilaments. The one exception was found in breast tissue
cultured in the presence of human chorionic gonadotropin, where the cells maintained differentiated characteristics, despite
the fact that they contained many lysosomes.
This work was supported by Grant R01 CA20764 and in part by Contract N01-CP-95640 awarded by the National Cancer Institute. 相似文献
11.
Campylobacter jejuni is an important food-borne pathogen. However, relatively little is understood regarding its pathogenesis, and research is hampered by the lack of a suitable model. Recently, a number of groups have developed assays to study the pathogenic mechanisms of C. jejuni using cell culture models. Here, we report the development of an ex vivo organ culture model, allowing for the maintenance of intestinal mucosal tissue, to permit more complex host-bacterium interactions to be studied. Ex vivo organ culture highlights the propensity for C. jejuni to adhere to mucosal tissue via the flagellum, either as discrete colonies or as multicellular units. 相似文献
12.
Effects of different concentrations of serum on cartilage growth in an organ culture system 总被引:1,自引:0,他引:1
R. Shurtz-Swirski D. Lewinson P. Shenzer M. Silbermann 《In vitro cellular & developmental biology. Plant》1989,25(11):995-999
Summary The purpose of the present study was to examine the effects of various concentrations of serum on the behavior of neonatal
condylar cartilage when cultured in an organ culture system. Mandibular condylar cartilages were obtained from newborn ICR
mice, of which the zone of undifferentiated chondroprogenitor cells along with a few layers of young cartilage cells were
cultivated at the medium-air interface. The incubation medium included fetal bovine serum at concentrations ranging from 0
to 10%, and the explants were kept in vitro up to 10 d. The serum-free medium maintained the chondrogenic expression, and
the overall size of the cartilagenous protion of the explants increased with the decrease of the concentrations of serum in
the medium. When explants were labeled with [3H]thymidine and were then processed for autoradiography, the peak of labeling was noticed at 48 h, a feature that recapitulated
itself in all cultures (73, 140, 175, 201, and 129 labeled cells per chondroprogenitor zone in explants grown in 0, 1, 2.5,
5, and 10%, respectively). It can be concluded that serum-free medium maintains the chondrogenic phenotype of condylar cartilage
in vitro.
This study was supported in part by a research grant from the Gesellschaft fur Biotechnologische Forschung mbH, Braunschweig-Stockheim,
Federal Republic of Germany. 相似文献
13.
14.
J. Feng A. H. Melcher D. M. Brunette H. K. Moe 《In vitro cellular & developmental biology. Plant》1977,13(2):91-99
Summary The method of Deutsch and Weeks was modified to provide a reliable and reasonably quick method for assaying the L-ascorbic
acid content of culture medium. The modified method was used to determine the decay of L-ascorbic acid under various conditions
of culture and the concentration of the vitamin in commercially prepared media. The half-life of L-ascorbic acid in a modified
New circulator gassed with 95% O2+5% CO2 was 1.5 hr.; and when gassed with 20% O2+5% CO2+75% N2, about 2 hr. In Petri dishes gassed with 20% O2+5% CO2+75% N2, the half-life of L-ascorbic acid was 0.9 hr. About 4% of the L-ascorbic acid was lost per day when medium was stored at
0°C and about 9% per day when stored at 5°C. When medium with an initial content of 300 μg per ml was stored at room temperature,
the half-life was found to be 15.5 hr. The L-ascorbic acid in five commercially available media, which contain the vitamin
in their formulations, was assayed immediately after their delivery to the laboratory. The values of L-ascorbic acid measured
in these media were in all cases far lower than prescribed. A continuous-flow organ culture system has been designed which
allows the provision of a relatively constant level of L-ascorbic acid to an explant by taking advantage of the slow oxidation
of L-ascorbic acid at 0°C. 相似文献
15.
Chemically-defined medium for growth and differentiation of mixed epithelial and connective tissues in organ culture 总被引:1,自引:0,他引:1
Gisele M. Hodges Anthony H. Melcher 《In vitro cellular & developmental biology. Plant》1976,12(6):450-459
The effect on tissue differentiation and growth in vitro of certain of the factors implicated in collagen synthesis (ascorbic
acid, α-ketoglutarate and oxygen) and the influence of hydrocortisone was studied using organ cultures of fetal mouse mandible
as a mixed epithelial and connective tissue system. Using serum-free Waymouth’s MB 752/1 chemically-defined medium, addition
of high levels of ascorbic acid (300 μg per ml), hydrocortisone (1 μg per ml) and oxygen (95%) enhanced differentiation in
a number of tissues, in particular skin and appendages, tooth germs and bone, while osteoid and dentine production were noticeably
promoted. It is suggested that an essential aspect of media design for organ culture involves the incorporation of collagen-promoting
factors to the in vitro environment particularly with regard to the controlling role implicated for collagen in a variety
of biological processes.
Some of the work reported here was undertaken while A. H. Melcher was a member of the Department of Dental Science, Royal
College of Surgeons of England, London, England. 相似文献
16.
Joseph T. M. Koumans Jean-yves Sire 《In vitro cellular & developmental biology. Animal》1996,32(10):612-626
Summary To develop a serum-free, chemically definedin vitro organ culture system enabling the study of epithelial-mesenchymal interactions in development and growth of fish dermal skeleton,
we investigatedin vitro continuation of scale regeneration in the cichlid fishHemichromis bimaculatus. The culture medium in our system is based on Leibovitz medium (L-15) supplemented with vitamin C, additional amino acids
and HEPES. With this basis medium, we examined the effects of all trans-retinoic acid, dexamethasone, and prostaglandin-E2
(PG-E2), factors known to exert an effect on development and growth of teeth and bone in mammalian culture systems, on thein vitro regeneration of scales. These effects were compared with those obtained by supplementation of the basis medium with newborn
and fetal calf serum. To evaluate our culture system, the medium that allowed to mimick in the best possible way thein vivo regeneration of scales (i.e., the basis medium plus dexamethasone and PG-E2) was also tested on thein vitro development of teeth in the same fish species.
Our serum-free, chemically defined organ culture system enablesin vitro development and growth of both scales and teeth. With this model culture system, it is possible to evaluate thein vitro effects of hormones, growth factors, and other substances on growth and development of dermal skeleton in fish. 相似文献
17.
18.
Daniel Acosta Elsie M. B. Sorensen David C. Anuforo David B. Mitchell Kenneth Ramos Kenneth S. Santone Mary Ann Smith 《In vitro cellular & developmental biology. Plant》1985,21(9):495-504
Summary A major goal of our laboratory has been the development of primary culture systems that retain differentiated fucntions and
responses characteristic of intact tissues in vivo. Specifically, we have developed cellular models of primary cultures of
rat heart, liver, and kidney cells to explore the mechanisms by which drugs or chemicals may be toxic to key organs of the
body and to develop new techniques by which xenobiotics may be evaluated or identified as potential toxicants to living systems.
The purpose of this paper is to describe our rationale and approach to the study of target organ toxicology with in vitro
cellular systems. 相似文献
19.
Several complex nutrient media were compared for their effectiveness in maintaining viable and functional mouse colon mucosa in organ culture. The order of superiority for preserving survival of normal tissues for 14 days was: Williams' Medium E > Morgan's 199 > CMRL-1066 > Waymouth's MB 752/1 > Eagle's MEM > Trowell's T8. The 3H-thymidine labeling index was highest in colon explants maintained in Morgan's 199 > Williams' Medium E > Waymouth's MB 752/1 > CMRL-1066 > Eagle's MEM. However, the very high labeling produced by Morgan's 199 medium was abnormal in comparison to in vivo levels. Supplementation with 1.0 µM dexamethasone almost always improved crypt survival and maintained normal DNA synthetic activity.Supported by National Cancer Institute contract No. 1-CP-75952 and grant No. CA-29602. 相似文献
20.
Abbott BD Buckalew AR Leffler KE 《Birth defects research. Part A, Clinical and molecular teratology》2005,73(6):447-454
BACKGROUND: 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is teratogenic in mice, producing cleft palate (CP). TCDD exposure disrupts expression of epidermal growth factor (EGF) receptor, EGF, and transforming growth factor-alpha (TGFalpha) in the palate and affects proliferation and differentiation of medial epithelial cells. EGF knockout embryos are less susceptible to the induction of CP by TCDD. This study used palate organ culture to examine the hypothesis that EGF enables a response to TCDD. METHODS: The midfacial tissues from wild-type (WT), EGF knockout, C57BL/6J, and TGFalpha knockout embryos were placed in organ culture on gestational day (GD) 12. Palatal explants were cultured for 4 days in serum-free Bigger's (BGJ) medium with 0.1% dimethyl sulfoxide (DMSO) or 1 x 10(-8) M TCDD with or without 2 ng of EGF/ml, 1 or 2 ng of TGFalpha/ml. Effects on palatal fusion were evaluated on day 4 of culture. EGF levels in explants and medium were determined using Luminex technology. RESULTS: In serum-free, control medium, palates from all of the strains fused. EGF knockout palates cultured with TCDD (no EGF) fused, but those cultured with TCDD + 2 ng of EGF/ml failed to fuse (p < 0.05 vs. control or TCDD without EGF). TGFalpha knockout palates failed to fuse when cultured with TCDD + 2 ng of TGFalpha/ml. EGF levels increased in tissue and accumulated in the medium after 24 hr of culture. CONCLUSIONS: This study demonstrated that providing EGF to the palates of EGF knockout mice restored the response to TCDD. These studies support the hypothesis that the mechanism for induction of CP by TCDD is mediated via the EGFR pathway. 相似文献