Comparison of a medium supplement - ‘Nu-Serum’ - and foetal calf serum in RPMI 1640 used for the murine allogeneic mixed lymphocyte reaction

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.

Download Full-text

‘Spontaneous’ sex reversal in organ cultures of the embryonic male gonad of the bird

Development ◽

10.1242/dev.31.3.611 ◽

1974 ◽

Vol 31 (3) ◽

pp. 611-620

Author(s):

Gregory F. Erickson

Keyword(s):

Germ Cells ◽

Primordial Germ Cells ◽

Calf Serum ◽

Sex Reversal ◽

Biologically Active ◽

Foetal Calf Serum ◽

Developmental Sequence ◽

Organ Cultures ◽

Male Gonad ◽

Ovarian Cortex

The left embryonic testis of the bird (4–8 days of incubation) was organ cultured in medium that contained 10% foetal calf serum. Under these conditions, the germinal epithelium (GE) of the 4-day gonad differentiates into an ovarian cortex and the male primordial germ cells (PGCs) complete a developmental sequence similar to normal oocytes, i.e. they divide mitotically, develop a Balbiani body, divide synchronously in groups of two, four, and eight germ cells, and some enter pre-leptotene. No medullary tissue develops in the 4-day explants. The pieces of 6- and 8-day gonad differentiate into true ovotestes in which the GE develops into a cortex and the medulla develops into seminiferous cords. The PGCs in the cortex differentiate as oocytes and those in the seminiferous cords differentiate as spermatogonia. The possibility that biologically active oestrogens are present in the growth medium is discussed.

Download Full-text

Siplizumab, an Anti-CD2 Monoclonal Antibody, Induces a Unique Set of Immune Modulatory Effects Compared to Alemtuzumab and Rabbit Anti-Thymocyte Globulin In Vitro

Frontiers in Immunology ◽

10.3389/fimmu.2020.592553 ◽

2020 ◽

Vol 11 ◽

Author(s):

Christian Binder ◽

Felix Sellberg ◽

Filip Cvetkovski ◽

Erik Berglund ◽

David Berglund

Keyword(s):

Cell Proliferation ◽

T Cells ◽

T Cell ◽

T Cell Activation ◽

Cell Activation ◽

Mixed Lymphocyte Reaction ◽

T Cell Proliferation ◽

Allogeneic Mixed Lymphocyte Reaction ◽

Dependent Cell

Antibodies are commonly used in organ transplant induction therapy and to treat autoimmune disorders. The effects of some biologics on the human immune system remain incompletely characterized and a deeper understanding of their mechanisms of action may provide useful insights for their clinical application. The goal of this study was to contrast the mechanistic properties of siplizumab with Alemtuzumab and rabbit Anti-Thymocyte Globulin (rATG). Mechanistic assay systems investigating antibody-dependent cell-mediated cytotoxicity, antibody-dependent cell phagocytosis and complement-dependent cytotoxicity were used to characterize siplizumab. Further, functional effects of siplizumab, Alemtuzumab, and rATG were investigated in allogeneic mixed lymphocyte reaction. Changes in T cell activation, T cell proliferation and frequency of naïve T cells, memory T cells and regulatory T cells induced by siplizumab, Alemtuzumab and rATG in allogeneic mixed lymphocyte reaction were assessed via flow cytometry. Siplizumab depleted T cells, decreased T cell activation, inhibited T cell proliferation and enriched naïve and bona fide regulatory T cells. Neither Alemtuzumab nor rATG induced the same combination of functional effects. The results presented in this study should be used for further in vitro and in vivo investigations that guide the clinical use of immune modulatory biologics.

Download Full-text

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

Cytotechnology ◽

10.1007/bf00749940 ◽

1993 ◽

Vol 13 (2) ◽

pp. 133-141 ◽

Cited By ~ 4

Author(s):

Bo Damgaard ◽

Anna Sanfeliu ◽

Jordi Joan Cair� ◽

Carles Casas ◽

Carles Sol� ◽

...

Keyword(s):

Serum Concentration ◽

Cell Line ◽

Calf Serum ◽

Foetal Calf Serum

Download Full-text

Molecular analysis of bovine viral diarrhoea virus isolates from South Africa

Onderstepoort Journal of Veterinary Research ◽

10.4102/ojvr.v70i4.292 ◽

2003 ◽

Vol 70 (4) ◽

Cited By ~ 6

Author(s):

N. Kabongo ◽

C. Baule ◽

M. Van Vuuren

Keyword(s):

South Africa ◽

Southern Africa ◽

Calf Serum ◽

Bovine Viral Diarrhoea Virus ◽

Border Disease Virus ◽

Foetal Calf Serum ◽

Type I ◽

Bovine Viral Diarrhoea ◽

Border Disease ◽

Diarrhoea Virus

The presence of bovine viral diarrhoea virus in South Africa has been confirmed by several serological surveys. However, little is known about its biological properties. Twenty five isolates obtained by isolation in tissue culture and detected by means of the antigen capture ELISA from clinically sick cattle and from foetal calf serum in South Africa were characterized on the basis of analysis of the 5' non-translated (NTR) region of the genome. A reverse-transcription polymerase chain reaction (RT-PCR) was used to amplify specific sequences from the 5'NTR of the genome. The oligonucleotide primers corresponding to positions 105-125 and 399-378, respectively, in the sequence of BVDV strain NADL were used to generate the PCR products. Both strands were sequenced directly with these primers and fluorescence-labelled dideoxynucleotides in an automated nucleic acid sequencer. Reference strains of pestiviruses [(BVDV type I, BVDV type II, border disease virus (BDV) and hog cholera virus (HCV)] and isolates from a previous investigation on BVDV in southern Africa were included for comparative purposes. All the BVDV strains obtained during this study belong to subgroups of BVDV genotype I. No association could be demonstrated between the geographic origin of the isolates. A number of isolates formed another branch separate from the existing branches Ia, Ib and Ic. These findings suggest that extensive genetic diversity can be found within BVDV type I isolates from southern Africa. Isolates that group with the classical BVDV type I strains, particularly of American origin, coexist with variants that appear to represent a local genetic pool and or variants evolving from the classical strains.

Download Full-text