STUDIES ON TISSUE CULTURE OF EQUINE OVARIAN CELL TYPES: EFFECT OF GONADOTROPHINS AND STAGE OF CYCLE ON STEROIDOGENESIS

1969 ◽  
Vol 43 (3) ◽  
pp. 415-425 ◽  
Author(s):  
CORNELIA P. CHANNING
Keyword(s):  
Tissue Culture ◽  
Cell Cultures ◽  
Granulosa Cells ◽  
Horse Serum ◽  
Cell Types ◽  
Phase Cell ◽  
Medium Change ◽  
Midluteal Phase

SUMMARY Granulosa cells were harvested from follicles of mares at various stages of the oestrous cycle and maintained in a tissue culture medium containing 15% horse serum, 30% medium '199' and 55% Hanks's solution. Between days 4 and 10 of culture the granulosa cells harvested from small follicles (1–2 cm. diam.) of mares in the midluteal phase of the cycle secreted an average of 0·36 pg. progesterone/cell/day. Cells harvested from large follicles of mares in the late and/or early oestrous stage of the cycle secreted an average of 29·5 pg. progesterone cell/day; the cells harvested from the large vascular follicles found at oestrus secreted an average of 173 pg./cell/day. The small, poorly vascularized follicles found adjacent to the large vascular follicles of mares in oestrus yielded cells which secreted less progesterone than those from the larger follicles. Addition of 5 to 10 i.u. human chorionic gonadotrophin (HCG)/ml. at each medium change (every 2–3 days) or for the first 4 days of culture brought about a marked stimulation of progesterone secretion in cultures of ' mid-luteal phase' cells which was maximal after 4 to 7 days. Pregnenolone was converted primarily to progesterone, 20α-hydroxypregn-4-en-3-one and 17-hydroxyprogesterone; the metabolism was not significantly altered by the addition of a mixture of 10 i.u. HCG plus 10 i.u. pregnant mare serum gonadotrophin (PMSG). Cells harvested from mares in oestrus converted pregnenolone to progesterone in a higher yield compared with cells harvested from mares in the midluteal phase of the cycle. Addition of 10 i.u. HCG/ml. or PMSG plus HCG (10 i.u. each/ml.) stimulated aromatization of testosterone by 'midluteal phase' cultures but not by 'oestrous phase' cell cultures. These results demonstrate that the in vivo environment as well as the in vitro conditions influence the steroidogenic activity of equine granulosa cell cultures.

In vitro interactions between epithelial cells and Gyrodactylus derjavini

2000 ◽  
Vol 74 (3) ◽  
pp. 203-208 ◽  
Author(s):  
K. Buchmann ◽  
C.V. Nielsen ◽  
J. Bresciani
Keyword(s):  
Tissue Culture ◽  
Epithelial Cells ◽  
Cell Cultures ◽  
Culture Medium ◽  
Epithelioma Papulosum Cyprini ◽  
Skin Responses ◽  
Enzyme Reactivity ◽  
In Vitro Interactions

AbstractSkin responses of fish to various parasites have been shown to involve various immunologically competent cells producing factors which guide the reactions of epithelial cells. However, the present study has demonstrated that a monoculture of epithelial cells has the ability to encapsulate and partially degrade ectoparasites without involvement of leukocytes. The ectoparasitic monogeneanGyrodactylus derjavini was kept on a monolayer of Epithelioma Papulosum Cyprini (EPC) cells in 24-well multidishes supplied with tissue culture medium. Gyrodactylus derjavini did not reproduce but survived an incubation period of up to139 h in the system. Due to sterile conditions, dead gyrodactylids were not subjected to microbial degradation and remained intact for several weeks. However, at 40 days G. derjavini was overgrown by EPC-cells and became partly degraded during the following 15 days. Analysis of enzyme reactivity in EPC-cells showed reactions for ten enzymes including esterases, amidases, phosphatases and phosphohydrolases. No marked differences for the ten enzymes between cell cultures with and without the ectoparasites were found but it cannot be excluded that some of these enzymes took part in parasite degradation. The study showed the in vitro capability of epithelial cells to interact, encapsulate and degrade G. derjavini without the involvement of leukocytes. This response probably is non-specific and will not exclude that various immunocompetent cells and their products normally optimize and accelerate elimination of invading parasites in vivo.

Immune responses to hapten conjugates in vitro

1975 ◽  
Vol 8 (4) ◽  
pp. 507-522
Author(s):  
Sirkka Kontiainen ◽  
O. Mäkelä ◽  
M. Hurme
Keyword(s):  
Tissue Culture ◽  
Immune Responses ◽  
Cell Types ◽  
Antibody Responses ◽  
Tissue Cultures ◽  
Cell Type ◽  
Animal Body ◽  
Do So

Several functions of the animal body can take place in cell or tissue cultures with almost unreduced efficiency and precision. Functions, where only one cell type is involved, often do so, but also some differentiation steps where interactions between two or more cell types are clearly needed can take place in tissue culture (Saxén et al. 1968).Most immune responses require collaboration between two or more cell types (Claman, Chaperon & Triplett, 1966; Miller & Mitchell, 1968; Feldmann & Nossal 1972c). Some of them can be easily induced in vitro but others cannot. Even when antibody responses can be induced in vitro their intensity varies a great deal. With some antigens and under some circumstances a response in vitro can be nearly as strong as one in vivo. A crude comparison can be derived from responses in vitro and in vivo to the same antigen, conjugate of hapten NIP and pneumococcal polysaccharide type III (NIP-SIll, Nakamura, Ray & Mäkelä, 1973).

scholarly journals STUDIES ON ANTIBODY FORMATION BY PERITONEAL EXUDATE CELLS IN VITRO

1960 ◽  
Vol 111 (4) ◽  
pp. 573-600 ◽  
Author(s):  
John M. McKenna ◽  
Kingsley M. Stevens
Keyword(s):  
Tissue Culture ◽  
Cell Types ◽  
Peritoneal Exudate ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Peritoneal Exudate Cells ◽  
Carbon Particles ◽  
Secondary Systems

Cells from peritoneal exudates of rabbits sacrificed 3 days after an intraperitoneal injection of sterile mineral oil were grown in tissue cultures in medium 199 (75 per cent); normal rabbit serum (25 per cent). Antibody produced by the cells was assayed by an hemagglutination technique in which the antigens used were adsorbed to formalinized tanned sheep erythrocytes. These sensitized cells agglutinate in the presence of antibody specific to the adsorbed antigen. It has been demonstrated that: Peritoneal exudate cells produced hemagglutinating antibody to bovine gamma globulin (BGG) in a replicating tissue culture system for approximately 3 weeks when taken from animals given either primary or secondary injections of BGG. The mean hemagglutinating titer was 30 for the primary and 32 for the secondary systems. Since the other cell types did not persist, it is felt that monocytes were responsible for these results. Monocytes taken from normal rabbits and exposed to either BGG or egg albumen (EA) in vitro produced titers of 28 for about 2 weeks. Monocytes taken from rabbits given hyperimmunizing injections of BGG produced titers of 147 for about 1 week. Endotoxin from Salmonella typhosa caused the monocytes to form antibody as if they had been taken from hyperimmunized rabbits. This was true both when the antigen was given in vivo together with the endotoxin as well as when the cells were exposed to antigen in vitro. The titers were 223 and 97, respectively. Neither freshly harvested nor cultured monocytes were phagocytic for carbon particles or bacteria in vitro. Monocytes in tissue culture appeared to assume the morphology of fibroblasts, but did not stain with the characteristics of fibroblasts. The morphologic changes and staining characteristics of monocytes in tissue culture have been described. The implications of these findings have been discussed and an attempt made to integrate them into general biological theory.

scholarly journals Fangchinoline Inhibits Human Esophageal Cancer by Transactivating ATF4 to Trigger Both Noxa-Dependent Intrinsic and DR5-Dependent Extrinsic Apoptosis

2021 ◽  
Vol 11 ◽  
Author(s):  
Yunjing Zhang ◽  
Shiwen Wang ◽  
Yukun Chen ◽  
Junqian Zhang ◽  
Jing Yang ◽  
...  
Keyword(s):  
Human Cancer ◽  
Cell Types ◽  
Underlying Mechanism ◽  
Phase Cell ◽  
Cycle Arrest ◽  
Extrinsic Apoptosis ◽  
Human Esophageal Cancer ◽  
First Time

Esophageal squamous cell carcinoma (ESCC) is a recalcitrant cancer. The Chinese herbal monomer fangchinoline (FCL) has been reported to have anti-tumor activity in several human cancer cell types. However, the therapeutic efficacy and underlying mechanism on ESCC remain to be elucidated. In the present study, for the first time, we demonstrated that FCL significantly suppressed the growth of ESCC both in vitro and in vivo. Mechanistic studies revealed that FCL-induced G1 phase cell-cycle arrest in ESCC which is dependent on p21 and p27. Moreover, we found that FCL coordinatively triggered Noxa-dependent intrinsic apoptosis and DR5-dependent extrinsic apoptosis by transactivating ATF4, which is a novel mechanism. Our findings elucidated the tumor-suppressive efficacy and mechanisms of FCL and demonstrated FCL is a potential anti-ESCC agent.

TISSUE CULTURE OF EQUINE OVARIAN CELL TYPES: CULTURE METHODS AND MORPHOLOGY

1969 ◽  
Vol 43 (3) ◽  
pp. 381-NP ◽  
Author(s):  
CORNELIA P. CHANNING
Keyword(s):  
Tissue Culture ◽  
Cell Division ◽  
Granulosa Cells ◽  
Lipid Droplets ◽  
Horse Serum ◽  
Cell Types ◽  
Culture Methods ◽  
Ovarian Cell ◽  
Chromatin Material ◽  
Fibroblastic Cells

SUMMARY Equine granulosa and other cell types have been cultured for up to 70 days in a medium containing 15% horse serum, 30% medium '199' and 55% Hanks's solution. Granulosa cells harvested from large (3–6 cm. in diameter) vascular follicles of mares in oestrus grew in epithelioid colonies. Cell division and hypertrophy lasted for 7 days with the greatest amount of hypertrophy occurring during the first 3–4 days in culture. Within 3 days the cytoplasm increased in size and acquired eosinophilic granules and lipid droplets; nuclei acquired distinct chromatin material and nucleoli. Cells harvested from small follicles grew in fibroblastic stellate colonies and underwent hypertrophy and hyperplasia but did not show any of the cytoplasmic characteristics of the cultures of granulosa cells obtained from the large follicles. Heterogeneous fibroblastic cells grew from explants of stromal and thecal tissue from days 5 to 20 of culture.

INFLUENCE OF 16-ARYLOXYPROSTAGLANDINS ON THE PRODUCTION OF PROGESTERONE BY HUMAN GRANULOSA CELLS IN VITRO

1976 ◽  
Vol 71 (2) ◽  
pp. 259-263 ◽  
Author(s):  
K. M. HENDERSON
Keyword(s):  
Tissue Culture ◽  
Adenylate Cyclase ◽  
Granulosa Cells ◽  
Farm Animals ◽  
Human Granulosa Cells ◽  
Progesterone Production ◽  
Porcine Granulosa Cells ◽  
Prostaglandin F

SUMMARY 16-Aryloxy analogues of prostaglandin F2α(PGF2α) are potent luteolysins in laboratory and farm animals. When their effect on progesterone production by luteinized human granulosa cells in tissue culture was investigated inhibition of both basal and gonadotrophin-stimulated progesterone production was observed, so revealing characteristics expected of potential human luteolysins. The analogues were, however, unable to inhibit progesterone production stimulated by PGE2, suggesting that like PGF2α these compounds may act by specifically blocking LH-activated adenylate cyclase. The 16-aryloxyprostaglandins similarly inhibited progesterone production by porcine granulosa cells, so that the effects observed with the 16-aryloxyprostaglandins in vitro may be indicative of their potential in vivo.

scholarly journals Simplified Bioprinting-Based 3D Cell Culture Infection Models for Virus Detection

Viruses ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 1298
Author(s):  
Robert Koban ◽  
Tobias Lam ◽  
Franziska Schwarz ◽  
Lutz Kloke ◽  
Silvio Bürge ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Cultures ◽  
Virus Detection ◽  
3D Cell Culture ◽  
Cell Types ◽  
Promising Alternative ◽  
Host Interactions ◽  
Infection Models

Studies of virus–host interactions in vitro may be hindered by biological characteristics of conventional monolayer cell cultures that differ from in vivo infection. Three-dimensional (3D) cell cultures show more in vivo-like characteristics and may represent a promising alternative for characterisation of infections. In this study, we established easy-to-handle cell culture platforms based on bioprinted 3D matrices for virus detection and characterisation. Different cell types were cultivated on these matrices and characterised for tissue-like growth characteristics regarding cell morphology and polarisation. Cells developed an in vivo-like morphology and long-term cultivation was possible on the matrices. Cell cultures were infected with viruses which differed in host range, tissue tropism, cytopathogenicity, and genomic organisation and virus morphology. Infections were characterised on molecular and imaging level. The transparent matrix substance allowed easy optical monitoring of cells and infection even via live-cell microscopy. In conclusion, we established an enhanced, standardised, easy-to-handle bioprinted 3D-cell culture system. The infection models are suitable for sensitive monitoring and characterisation of virus–host interactions and replication of different viruses under physiologically relevant conditions. Individual cell culture models can further be combined to a multicellular array. This generates a potent diagnostic tool for propagation and characterisation of viruses from diagnostic samples.

STUDIES ON TISSUE CULTURE OF EQUINE OVARIAN CELL TYPES: PATHWAYS OF STEROIDOGENESIS

1969 ◽  
Vol 43 (3) ◽  
pp. 403-414 ◽  
Author(s):  
CORNELIA P. CHANNING
Keyword(s):  
Tissue Culture ◽  
Granulosa Cell ◽  
Cell Cultures ◽  
Granulosa Cells ◽  
Sodium Acetate ◽  
Steroid Hormone ◽  
Cell Types ◽  
Ovarian Cell ◽  
Luteal Tissue ◽  
17 Hydroxyprogesterone

SUMMARY Equine granulosa cell cultures were incubated with various labelled steroid hormone precursors, and the products of these incubations were purified and characterized by a combination of paper chromatography, derivative formation and recrystallization. Sodium acetate, cholesterol and pregnenolone were converted mainly to progesterone in addition to significant amounts of 20α-hydroxypregn-4-en-3-one and 17-hydroxyprogesterone plus small amounts of dehydroepiandrosterone, androstenedione and oestradiol-17β. Both androstenedione and testosterone were converted to oestrone and oestradiol-17β. Testosterone was converted to androstenedione but the converse did not apply. These experiments demonstrated that equine granulosa cells in culture contain all the enzymes for converting acetate to progesterone and oestradiol-17β and that the pathways are similar to those of luteal tissue.

scholarly journals The Plasma [Kynurenine]/[Tryptophan] Ratio and Indoleamine 2,3-Dioxygenase: Time for Appraisal

2019 ◽  
Vol 12 ◽  
pp. 117864691986897 ◽  
Author(s):  
Abdulla A-B Badawy ◽  
Gilles Guillemin
Keyword(s):  
Extracellular Matrix ◽  
Cell Cultures ◽  
Cell Types ◽  
Future Studies ◽  
Isolated Organs ◽  
Indoleamine 2,3 Dioxygenase ◽  
Multiple Cell ◽  
Complex Structural

The plasma kynurenine to tryptophan ([Kyn]/[Trp]) ratio is frequently used to express or reflect the activity of the extrahepatic Trp-degrading enzyme indoleamine 2,3-dioxygenase (IDO). This ratio is increasingly used instead of measurement of IDO activity, which is often low or undetectable in immune and other cells under basal conditions, but is greatly enhanced after immune activation. The use of this ratio is valid in in vitro studies, eg, in cell cultures or isolated organs, but its ‘blanket’ use in in vivo situations is not, because of modulating factors, such as supply of nutrients; the presence of multiple cell types; complex structural and functional tissue arrangements; the extracellular matrix; and hormonal, cytokine, and paracrine interactions. Determinants other than IDO may therefore be involved in vivo. These are hepatic tryptophan 2,3-dioxygenase (TDO) activity and the flux of plasma-free Trp down the Kyn pathway. In addition, conditions leading to accumulation of Kyn, eg, inhibition of activities of Kyn monooxygenase and kynureninase, could lead to elevation of the aforementioned ratio. In this review, the origin of use of this ratio will be discussed, variations in extent of its elevation will be described, evidence against its indiscriminate use will be presented, and examining determinants other than IDO activity and their correlates will be proposed for future studies.

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