Cultivation of Blastocrithidia triatomae (Trypanosomatidae) on a cell line of its host Triatoma infestans (Reduviidae)

Parasitology ◽  
1989 ◽  
Vol 98 (3) ◽  
pp. 387-393 ◽  
Author(s):  
Dagmar Reduth ◽  
G. A. Schaub ◽  
Mary Pudney
Keyword(s):  
Cell Line ◽  
Primary Cultures ◽  
Calf Serum ◽  
Host Cells ◽  
Triatoma Infestans ◽  
Foetal Calf Serum ◽  
Tryptose Phosphate Broth ◽  
A Cell ◽  
Vigorous Growth

SUMMARYBlastocrithidia triatomae parasitizes vectors of Chagas' disease and is very difficult to cultivate in conventional media. However, co-cultivation with a cell line of its host Triatoma infestans (TI-32; in Schneider's Drosophila medium supplemented with 20% foetal calf serum and 10% tryptose phosphate broth) led to vigorous growth at 24 or 28 °C without an adaptation phase. More than 60 primary cultures were initiated successfully without any failures. Subcultures could be started immediately at weekly intervals. The doubling time was similar in both the primary cultures (41 h) and the 33rd subculture (39 h). The importance of the reduviid cells for B. triatomae became clear after removal of the insect cells, when multiplication of epimastigotes stopped and mainly cysts were formed. Cysts produced in vitro were infective for reduviids. Scanning electron microscopy showed that B. triatomae attached to the host cells, inserted its flagellum into them and destroyed them.

Endocrine and mucous differentiation by a cloned human rectal adenocarcinoma cell line (HRA-19) in vitro: inhibition by TGF-beta 1

1994 ◽  
Vol 107 (4) ◽  
pp. 1041-1046
Author(s):  
S.C. Kirkland ◽  
K. Henderson
Keyword(s):  
Cell Line ◽  
Endocrine Cells ◽  
Calf Serum ◽  
Rectal Adenocarcinoma ◽  
Model Systems ◽  
Foetal Calf Serum ◽  
Mucous Cells ◽  
Extrinsic Factors ◽  
Colorectal Epithelium

Colorectal epithelium is composed of absorptive, mucous and endocrine cells, all of which are considered to arise from a common stem cell located in the crypt base. However, the factors controlling the commitment to differentiate are poorly understood. This is partly due to the lack of in vitro model systems for the study of differentiation in colorectal epithelium. The HRA-19 cell line, established from a human rectal adenocarcinoma, has been shown to have multipotential characteristics with cloned HRA-19 cells able to differentiate into absorptive, mucous and endocrine cells when grown as xenografts. The lack of such differentiated cells in HRA-19 monolayers in vitro suggests that differentiation is controlled by extracellular matrix, stromal cells and/or soluble factors. Such observations show that differentiation in HRA-19 cells can be controlled by extrinsic factors and therefore provide a model system for studying control of differentiation in colorectal epithelium. Unfortunately, the restriction of differentiation to xenografts of the cell line limits the degree to which this differentiation can be manipulated. In this study, the possibility that HRA-19 cells could be induced to differentiate in vitro under appropriate conditions has been investigated. Endocrine and mucous cells were identified by immunocytochemistry with differentiation-related antibodies and histology of monolayers. Preconfluent HRA-19 cells grown in 10% foetal calf serum formed a well polarised monolayer with apical tight junctions and sparse microvilli, but cells with mucous or endocrine phenotypes were only very occasionally observed. However, endocrine and mucous cells could reproducibly be demonstrated in postconfluent monolayers grown in 1% foetal calf serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Optimization of culture conditions for the maintenance of Onchocerca gutturosa adult worms in vitro

1986 ◽  
Vol 60 (4) ◽  
pp. 323-330 ◽  
Author(s):  
Simon Townson ◽  
C. Connelly ◽  
R. Muller
Keyword(s):  
Calf Serum ◽  
Culture Conditions ◽  
Foetal Calf Serum ◽  
Free System ◽  
Gas Phases ◽  
Mdbk Cells ◽  
A Cell ◽  
Series Of Experiments

ABSTRACTA series of experiments examined the effects of various media, serum supplements, gas phases and the incorporation of mammalian cell feeder layers on the survival on Onchocerca gutturosa adult worms in vitro. The survival of male worms was poor in all media tested that were not supplemented with inactivated foetal calf serum (IFCS), with improved but variable survival in media supplemented with 10–30% IFCS. Using a cell-free system in an atmosphere of 5% CO2in air, good results were obtained in medium NCTC 135 + 10% IFCS (median survival time 39 days, range 25–41). Marginally better survival was obtained with the same medium in an atmosphere of 95% N2/5% CO2 (medium 45 days, range 25–56)and with a 1:1 mixture of media NCTC 135 and IMDM+10% IFCS(medium 38 days, range 38–51). Survival was enhanced in culture systems which incorporated bovine kindney (MDBK) cells, bovine trachea (EBTR) cells and monkey kidney (LLCMK2) cells. Exceptionally long survival was obtained using medium MEM+10% IFCS+LLCMK2 cells under a gas phase of 5% CO2in air, in which male worms survived from approximately 6 to over 7 months. Under similar conditions, female worms were also maintained for periods of up to 6 months and 5 out of 18 specimens released microfilariae into the culture system. The long-term culture described in this study will be useful for basic biochemical, chemotherapeutic and immunological studies in vitro.

scholarly journals PO02CHARACTERISATION OF GLIOBLASTOMA METABOLISM IN-VITRO: A COMPARISON BETWEEN A CELL LINE AND PRIMARY CULTURES

2015 ◽  
Vol 17 (suppl 8) ◽  
pp. viii1.2-viii1
Author(s):  
M. Waqar ◽  
R. Richards ◽  
V. See
Keyword(s):  
Cell Line ◽  
Primary Cultures ◽  
A Cell

Drug activity against Onchocerca gutturosa males in vitro: a model for chemotherapeutic research on onchocerciasis

1987 ◽  
Vol 61 (4) ◽  
pp. 271-281 ◽  
Author(s):  
Simon Townson ◽  
C. Connelly ◽  
A. Dobinson ◽  
R. Muller
Keyword(s):  
Culture System ◽  
Serum Proteins ◽  
Good Condition ◽  
Calf Serum ◽  
New Drugs ◽  
Foetal Calf Serum ◽  
Partial Recovery ◽  
Feeder Cells ◽  
Compound Identification

ABSTRACTAn in vitro system for chemotherapeutic research using adult male Onchocerca gutturosa has been developed as a model for O. volvulus. Using a culture system consisting of medium MEM+10% heat inactivated foetal calf serum (IFCS)+LLCMK2 (monkey kidney) feeder cells in an atmosphere of 5% CO2 in air, we examined the effects of a range of antiparasitic drugs on worm motility. Ivermectin, levamisole, furapyrimidone, Mel W, chloroquine, metrifonate, flubendazole, amoscanate and the Ciba-Geigy compounds CGP 6140, CGP 20′376 and CGI 17658 either immobilized or significantly reduced motility levels at a concentration of 5x10−5M or less within a 7-day period. Worms were affected at very low concentrations by ivermectin (effective conc. to reduce motility levels to 50% of controls, 3.14x10−8M), levamisole (7.95x10−8M), CGP 6140 (8.87x10−9M) and CGP 20′376 (2.78x10−8M). Difficulties were experienced in accurately repeating the immotile endpoint for levamisole due to an inconsistent partial recovery of motility. Over a 7-day period diethylcarbamazine had little effect on motility levels, while suramin caused a slight increase in activity compared to controls at some timepoints. Subsequent experiments demonstrated some differences in drug efficacy depending on the presence or absence of serum and feeder cells in the culture system probably because of drug avidly binding to serum proteins. However, serum and cells were found to be essential ingredients of the culture system to maintain worms in good condition, indicating that new drugs should be evaluated both in the presence and absence of serum and cells. Comparisons were made between the responses of O. gutturosa and Brugia pahangi to certain drugs and these species were found to significantly differ in their sensitivities to ivermectin and a novel compound (Wellcome), indicating that Onchocerca parasites should be used wherever possible for compound identification and development intended for the treatment of onchocerciasis. The in vitro system described here, using male O. gutturosa, provides a basis for further research and a practical alternative to O. volvulus.

Production of Human Chorionic Gonadotropin In Vitro by a Cell Line Derived From a Carcinoma of the Lung2

1973 ◽  
Vol 50 (3) ◽  
pp. 669-674 ◽  
Author(s):  
Alan S. Robson ◽  
Saul W. Rosen ◽  
Armen H. Tashjian ◽  
Bruce D. Weintraub
Keyword(s):  
Cell Line ◽  
Chorionic Gonadotropin ◽  
Human Chorionic Gonadotropin ◽  
A Cell

Substitution of transferrin by FeCl3 in the development of a low foetal calf serum concentration medium for KB-26.5 hybridoma cell line

1993 ◽  
Vol 13 (2) ◽  
pp. 133-141 ◽  
Author(s):  
Bo Damgaard ◽  
Anna Sanfeliu ◽  
Jordi Joan Cair� ◽  
Carles Casas ◽  
Carles Sol� ◽  
...  
Keyword(s):  
Serum Concentration ◽  
Cell Line ◽  
Calf Serum ◽  
Foetal Calf Serum

scholarly journals The effect of strontium ranelate on titanium particle-induced periprosthetic osteolysis regulated by WNT/β-catenin signaling in vivo and in vitro

2021 ◽  
Vol 41 (1) ◽  
Author(s):  
Bolun Wang ◽  
Haohui Guo ◽  
Tianxiang Geng ◽  
Kening Sun ◽  
Liang Zhang ◽  
...  
Keyword(s):  
Bone Resorption ◽  
Aseptic Loosening ◽  
Cell Line ◽  
Conditioned Medium ◽  
Strontium Ranelate ◽  
Cell Model ◽  
Periprosthetic Osteolysis ◽  
A Cell

Abstract Aseptic loosening following periprosthetic osteolysis is the primary complication that limits the lifetime of total joint arthroplasty (TJA). The wear particles trigger a chronic inflammation response in the periprosthetic tissue and turn over the bone balance to bone resorption. The present study aimed to investigate the possible effect and mechanism of strontium ranelate (SR), a clinically safe drug for osteoporosis, on particle-induced periprosthetic osteolysis. Thirty-six female C57BL/6j mice underwent tibial Ti-nail implantation to establish an animal model of aseptic loosening. After 12 weeks, micro-CT results showed that strontium ranelate could inhibit periprosthetic bone resorption. In vitro, Ti particles were used to stimulate RAW264.7 cell line to collect conditioned medium, and co-culture MC3T3-E1 cell line with conditioned medium to establish a cell model of aseptic loosening. The results of alkaline phosphatase (ALP) detection, immunofluorescence, and flow cytometry demonstrated that strontium ranelate could regulate the expression of OPG/RANKL, promote differentiation and mineralization, and inhibit apoptosis in osteoblasts. Moreover, we revealed that SR’s exerted its therapeutic effect by down-regulating sclerostin, thereby activating the Wnt/β-catenin signal pathway. Therefore, this research suggests that strontium ranelate could be a potential drug for the prevention and treatment of particle-induced aseptic loosening post-TJA.

scholarly journals Modelling Bovine Granuloma Formation In Vitro upon Infection with Mycobacterium Avium Subspecies Paratuberculosis

2019 ◽  
Vol 6 (4) ◽  
pp. 80 ◽  
Author(s):  
J. Hunter Rice ◽  
Margaret M. McDaniel ◽  
Alyson Holland ◽  
Shigetoshi Eda
Keyword(s):  
Mycobacterium Avium ◽  
Mycobacterium Avium Subspecies Paratuberculosis ◽  
Clinical Signs ◽  
Host Cells ◽  
Economic Losses ◽  
Multiple Parameters ◽  
Cell Mediated Immune Response ◽  
A Cell ◽  
Vitro Model

Mycobacterium avium subspecies paratuberculosis (Map) causes chronic granulomatous disease in cattle and ruminant livestock, causing substantial economic losses. Current vaccines delay clinical signs but cannot train the immune system to fully eradicate latent Map. During latency, Map uses host defenses, cage-like macrophage clusters called granuloma, as incubators for months or years. We used an in vitro model to investigate the early coordination of macrophages into granuloma upon Map infection over ten days. We found that at multiplicities of infection (MOI; Map:macrophages) of 1:2 and below, the macrophages readily form clusters and evolve pro-inflammatory cytokines in keeping with a cell-mediated immune response. At higher MOIs, viability of host macrophages is negatively impacted. At 1:4 MOI, we quantified viable Map in our model and confirmed that intracellular Map reproduced over the first five days of infection. Host cells expressed Type 1-specific cytokines, and Map-infected macrophages displayed reduced motility compared to Map-exposed, uninfected macrophages, suggesting an important role for uninfected macrophages in the early aggregative response. Reported is the first in vitro JD granuloma model capturing Map and macrophage viability, size distribution of resulting clusters, motility of monocyte-derived macrophages, and cytokine response during clustering, allowing quantitative analysis of multiple parameters of the Map-specific granulomatous response.

Maturation of porcine oocytes for 33 or 44 hours in vitro in the presence or absence of serum

1998 ◽  
Vol 1998 ◽  
pp. 180-180
Author(s):  
K. Rust ◽  
M.E. Staines ◽  
GJ. McCallum ◽  
N.S. Prathalingam ◽  
S.A. Edwards ◽  
...  
Keyword(s):  
Disease Transmission ◽  
Calf Serum ◽  
International Markets ◽  
Foetal Calf Serum ◽  
Maturation Time ◽  
Nuclear Maturation ◽  
Defined Media ◽  
Porcine Embryo ◽  
Disease Risks

Porcine embryo production in vitrois providing the impetus for the development of cryopreservation strategies aimed at welfare-friendly domestic and international marketing and movement of stock in a manner that minimises risks of disease transmission. In the context of disease risks, defined media, which avoid the use of serum and other biohazardous products, are likely to become essential in the production of embryos for international markets. In preparing for this situation, the present comparative study investigated in vitronuclear maturation of porcine oocytes in the presence of either foetal calf serum (FCS) or polyvinyl alcohol (PVA). In addition, the effect of restricting the maturation time to 33 rather than 44 hours was examined.

Sign in / Sign up

Export Citation Format

Share Document