scholarly journals Occurrence of a cytosolic neutral chitobiase activity involved in oligomannoside degradation: a study with Madin-Darby bovine kidney (MDBK) cells

1996 ◽  
Vol 313 (2) ◽  
pp. 597-602 ◽  
Author(s):  
René CACAN ◽  
Catherine DENGREMONT ◽  
Odette LABIAU ◽  
Daniel KMIÉCIK ◽  
Anne-Marie MIR ◽  
...  
Keyword(s):  
Cell Line ◽  
Mdbk Cell ◽  
Biological Models ◽  
Bovine Kidney ◽  
Streptolysin O ◽  
Mdbk Cells

Neutral oligomannosides possessing one GlcNAc (OS-Gn1) and two GlcNAc (Os-Gn2) at the reducing end have been reported to be released during the N-glycosylation process in various biological models. To investigate which enzyme is responsible for OS-Gn1 formation, we used the Madin-Darby bovine kidney (MDBK) cell line which exhibits neither lysosomal chitobiase nor endoglucosaminidase activities. However, these cells produced OS-Gn1 and we showed that a neutral chitobiase is responsible for the transformation of OS-Gn2 into OS-Gn1. Using streptolysin O-permeabilized MDBK cells, we demonstrated that this neutral chitobiase activity is located in the cytosolic compartment and is active on oligomannoside species released during the N-glycosylation process.

scholarly journals IN VITRO ASSESSMENT OF THE BIOLOGICAL ACTIVITY OF A NEW REGENERATIVE AGENT PREPARED FROM THE CONCENTRATE OF DEPROTEINIZED DERMAL LAYER OF PORCINE SKIN

2020 ◽  
pp. 12-22
Author(s):  
Natalia Bezdieniezhnykh ◽  
Aleksandra Lykhova ◽  
Hennadii Borschevskyi ◽  
Kateryna Dyakun ◽  
Ievgen Kruglov
Keyword(s):  
Biological Activity ◽  
Cell Line ◽  
Mammalian Cells ◽  
Therapeutic Agent ◽  
Biological Effects ◽  
Mdbk Cell ◽  
Porcine Skin ◽  
Bovine Kidney ◽  
Dermal Layer

Background. Presently, a prospective direction for the development of regenerative medicine in the world is the search for regulatory molecules and the identification of molecular targets to stimulate the body's endogenous regenerative potential. The concentrate of the deproteinized dermal layer of porcine skin (СDDLPS) is a new therapeutic agent with restorative properties, the action of which is directed on the induction of the self resources of cells. Aim. The assessment of the effect of СDDLPS on the proliferative activity of mammalian cells of different histogenesis in vitro. Materials and Methods. To determine the amino acid composition of the СDDLPS liquid chromatography and biochemical methods were used. The biological effects and mechanisms of action of the drug were investigated by cell culture and molecular biological methods. The research was carried out using stable cell lines: human keratinocytes (HaCaT cell line), porcine endothelial cells (PAE cell line), bovine kidney cells (MDBK cell line) and mouse fibroblasts (3T3A31 cell line). Results. The cells of the bovine kidney MDBK cell line were the most sensitive to the effect of the CDDLPS. Also, the obtained results suggest that, depending on the concentration, the drug not only stimulates cell proliferation by 10–30 %, but also significantly enhances biosynthetic processes in cells, in particular, protein synthesis by 20–40 %. Conclusions. CDDLPS is an effective and affordable therapeutic agent with restorative properties, the biological activity of which manifests itself in the activation of cell biosynthetic and proliferative potentials and is comparable to effects of some growth factors, in particular epidermal growth factor

scholarly journals Correction of genetic instability of the genome by fractional irradiation of MDBK cells

2020 ◽  
Vol 11 (2) ◽  
pp. 1879-1882
Author(s):  
Edie M. Plotnicova ◽  
Ramzi N. Nizamov ◽  
Ravil G. Fazliahmetov ◽  
Irina A. Arkharova ◽  
Almaz S. Saifullin ◽  
...  
Keyword(s):  
Cell Line ◽  
Standard Procedure ◽  
Test Dose ◽  
Mdbk Cell ◽  
Animal Origin ◽  
Federal State ◽  
State Budget ◽  
Logarithmic Growth Phase ◽  
Mdbk Cells ◽  
Fractional Irradiation

With respect to given contradictory data in radiobiology, that according to some authors, irradiation of cells of plant and animal origin initiates DNA changes and according to others, repeated irradiation of cells in small doses has a stimulating effect on metabolism without increasing the purity of mutations, we conducted the present study, with the aim to study the stability of the genome of MDBK cell line against a background of single and double (fractional) irradiation. The object of the study was a continuous MDBK cell line (Madin-Darby Bovire Kidney Cell, cattle, kidney), which was grown in medium 199, manufactured by Federal State Budget Institution FCTRB-ARRVI (Kazan) with 10% bovine serum and the addition of antibiotics (benzylpenicillin sodium salt, kanamycin at 100 units/ml) (streptomycin sulfate at 100 μg/ml) according to standard procedure. A monolayer culture of MDBK cells in the logarithmic growth phase (3-4 days after culture) served as contacting cells. As a result, a radio-modified cell line, called MDBK-2, was obtained. Moreover, in an unirradiated MDBK cell culture, the level of chromosomal breakdowns was in the range of 1.03-3.1% with respect to the total number of studied anaphases. Based on the obtained data, it can be concluded, that during prolonged cultivation of MDBK cells, irradiated in a small (adaptive) dose and re-irradiated in 3-minute higher dose (5.95 Gy), there is a significant decrease in the yield of aberrant cells, induced by a test dose of γ-rays.

scholarly journals Establishment of a Suspension MDBK Cell Line in Serum-Free Medium for Production of Bovine Alphaherpesvirus-1

Vaccines ◽  
2021 ◽  
Vol 9 (9) ◽  
pp. 1006
Author(s):  
Pengpeng Wang ◽  
Shulin Huang ◽  
Chengwu Hao ◽  
Zhanhui Wang ◽  
Haoran Zhao ◽  
...  
Keyword(s):  
Cell Line ◽  
Neutralizing Antibodies ◽  
Scale Up ◽  
Virus Titer ◽  
Mdbk Cell ◽  
Serum Free Medium ◽  
Free Medium ◽  
Suspension Cells ◽  
Serum Free ◽  
Mdbk Cells

The Madin–Darby bovine kidney (MDBK) cell line is currently used for the production of bovine alphaherpesvirus-1 (BoHV-1) vaccine. For the purpose of vaccine manufacturing, suspension cells are preferred over adherent ones due to simplified sub-cultivation and an easier scale-up process, both of which could significantly reduce production cost. This study aimed to establish a procedure for the culture of BoHV-1 in the suspended MDBK cell line in serum-free medium. We screened several commercially available serum-free media and chose ST503 for subsequent experiments. We successfully adapted the adherent MDBK cells to suspended growth in ST503 in the absence of serum. The maximum density of suspension-adapted MDBK cells could reach 2.5 × 107 cells/mL in ST503 medium with optimal conditions. The average size of suspension-adapted cells increased to 18 ± 1 µm from 16 ± 1 µm. Moreover, we examined tumorigenicity of the suspended cells and found no sign of tumorigenicity post adaptation. Next, we developed a protocol for the culture of BoHV-1 in the cell line described above and found that ultrasonic treatment could facilitate virus release and enhance virus yield by 11-fold, with the virus titer reaching 8.0 ± 0.2 log10TCID50/mL. Most importantly, the prototype inactivated BoHV-1 vaccine we generated using the suspension cultures of MDBK cells induced neutralizing antibodies to a titer comparable to that of the commercial inactivated BoHV-1 vaccine. Overall, we established and optimized a protocol for the production of inactivated BoHV-1 vaccine in MDBK cells adapted for suspension culture, which provides insights for future large-scale manufacturing of BoHV-1 vaccine.

scholarly journals Madin-Darby bovine kidney (MDBK) cells are a suitable cell line for the propagation and study of the bovine poxvirus lumpy skin disease virus

2020 ◽  
Vol 285 ◽  
pp. 113943
Author(s):  
Petra C. Fay ◽  
Charlotte G. Cook ◽  
Najith Wijesiriwardana ◽  
Gessica Tore ◽  
Loic Comtet ◽  
...  
Keyword(s):  
Cell Line ◽  
Disease Virus ◽  
Skin Disease ◽  
Lumpy Skin Disease ◽  
Lumpy Skin Disease Virus ◽  
Bovine Kidney ◽  
Mdbk Cells

 Evaluating large spontaneous deletions in a bovine cell line selected for bovine viral diarrhea virus resistance - MDBK reads mapping to regions in genome  not shared with CRIB cell line v1

2021 ◽  
Author(s):  
aspen.workman not provided ◽  
mike.heaton not provided ◽  
Dennis A. Webster ◽  
Gregory P Harhay ◽  
Tim Smith ◽  
...  
Keyword(s):  
Cell Line ◽  
Bovine Viral Diarrhea Virus ◽  
Virus Entry ◽  
Mdbk Cell ◽  
Genomic Variation ◽  
Host Factors ◽  
Bovine Viral Diarrhea ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

Bovine viral diarrhea virus (BVDV) entry into bovine cells involves attachment of virions to cellular receptors, internalization, and pH-dependent fusion with endosomal membranes. The primary host receptor for BVDV is CD46; however, the complete set of host factors required for virus entry is unknown. The Madin-Darby bovine kidney (MDBK) cell line is susceptible to BVDV infection, while a derivative cell line (CRIB) is resistant at the level of virus entry. We performed complete genome sequencing of each to identify genomic variation underlying the resistant phenotype with the aim of identifying host factors essential for BVDV entry. Three large compound deletions in the BVDV-resistant CRIB cell line were identified and predicted to disrupt the function or expression of the genesPTPN12,GRID2, andRABGAP1L. However, CRISPR/Cas9 mediated knockout of these genes, individually or in combination, in the parental MDBK cell line did not impact virus entry or replication. Therefore, resistance to BVDV in the CRIB cell line is not due to the apparent spontaneous loss ofPTPN12,GRID2, orRABGAP1Lgene function. Identifying the functional cause of BVDV resistance in the CRIB cell line may require more detailed comparisons of the genomes and epigenomes.

scholarly journals Approaches to probiotics properties testing using Caco-2 cells

2020 ◽  
Vol 175 ◽  
pp. 03024
Author(s):  
Viktor Chmykhalo ◽  
Peter Zolotukhin ◽  
Viktor Pakhomov ◽  
Aleksey Prutskov ◽  
Sabina Khairullina ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Line ◽  
Present Review ◽  
Biological Models ◽  
Intercellular Interactions ◽  
Wide Range

Adequate biological models are a prerequisite to screening and development of probiotic drugs. In the present study, Caco-2 cell line is reviewed as a model for analyzing properties of probiotics. This cell culture possesses all the characteristics necessary for evaluating the effects of probiotic drugs on a wide range of both intracellular processes and intercellular interactions. Informativeness of Caco-2 cells is in the focus of the present review.

Inhibition of the development of Eimeria tenella in cultured bovine kidney cells by a soluble factor produced by peripheral blood lymphocytes from immune chickens

Parasitology ◽  
1998 ◽  
Vol 117 (1) ◽  
pp. 39-47 ◽  
Author(s):  
J. M. BUMSTEAD ◽  
S. J. TOPHAM ◽  
F. M. TOMLEY
Keyword(s):  
Inhibitory Activity ◽  
Peripheral Blood ◽  
Culture Media ◽  
Peripheral Blood Lymphocytes ◽  
Eimeria Tenella ◽  
Bovine Kidney ◽  
Oral Infections ◽  
Blood Lymphocytes ◽  
Mdbk Cells

The intracellular development of Eimeria tenella sporozoites in in vitro cultured Madin–Darby Bovine Kidney (MDBK) cells was inhibited when parasite-infected MDBK cells were incubated with peripheral blood lymphocytes (PBL) from infected chickens. The inhibition mediated by PBL was quantified by [3H]uracil uptake and increased during the course of a series of oral infections of chickens with E. tenella. This was mirrored by the development of immunity in these birds, as assessed by counting the oocyst output following each re-infection. Similar levels of inhibition were observed using PBL from 3 inbred lines of chickens which differ in their relative susceptibility to infection with E. tenella, indicating that the genetic background of the host does not influence the production of this inhibitory activity. The inhibition could be transferred to freshly infected MDBK cells using supernatants prepared from parasite-infected monolayers incubated for 48 h with PBL from immune chickens. However, there was no inhibition using either supernatants from infected MDBK cells incubated with PBL from uninfected chickens, or supernatants from uninfected MDBK cells incubated with PBL from immune chickens. Experiments using Transwell plates showed that direct contact of PBL from immune birds with infected MDBK monolayers was not required to produce supernatants with inhibitory activity. Thus production of soluble inhibitory factor(s) by PBL from immune chickens can be specifically induced by soluble antigens present in the culture media of parasite-infected MDBK cells. These factors inhibit the intracellular development of sporozoites in in vitro culture.

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