Adaptation of an Indigenous Insect Cell Line DZNU-Bm-1 to Mitsuhashi and Maramorosch (MM) Culture Medium Supplemented with 3% Fetal Bovine Serum

2020 ◽  
Vol 13 (2) ◽  
pp. 842-847
Author(s):  
Syed Obaid Qureshi
Keyword(s):  
Cell Line ◽  
Insect Cell ◽  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Insect Cell Line ◽  
Fetal Bovine

Adaptation of an Aedes aegypti cell line to hemolymph-free culture medium

1969 ◽  
Vol 15 (10) ◽  
pp. 1197-1200 ◽  
Author(s):  
S. S. Sohi
Keyword(s):  
Aedes Aegypti ◽  
Bombyx Mori ◽  
Cell Line ◽  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Free Medium ◽  
Antheraea Pernyi ◽  
Fetal Bovine ◽  
In The Beginning

Aedes aegypti cells (Grace's line) growing in a medium containing 5% Antheraea pernyi hemolymph (APH) and 5% fetal bovine serum (FBS) were adapted to a hemolymph-free medium. The quantity of APH was gradually substituted with an equal quantity of FBS over a 2-month period. The cells grew very slowly in the hemolymph-free medium in the beginning. However, they became adapted to it in another 8 months; after that they grew as well as the cells in medium supplemented with hemolymph.Also, these cells grew as well in medium containing Bombyx mori hemolymph as in medium enriched with APH.

Adaptation of an insect cell line (Agallia constricta) in a mammalian cell culture medium

In Vitro ◽  
1973 ◽  
Vol 8 (5) ◽  
pp. 375-378 ◽  
Author(s):  
Arthur H. Intosh ◽  
K. Maramorosch ◽  
C. Rechtoris
Keyword(s):  
Cell Culture ◽  
Cell Line ◽  
Mammalian Cell ◽  
Insect Cell ◽  
Culture Medium ◽  
Mammalian Cell Culture ◽  
Insect Cell Line ◽  
Cell Culture Medium

Fetal bovine serum induces changes in fatty acid composition ofTrypanosoma cruziphosphoinositides

1995 ◽  
Vol 41 (10) ◽  
pp. 951-954 ◽  
Author(s):  
G. Racagni ◽  
M. G. de Lema ◽  
G. Hernández ◽  
E. E. Machado-Domenech
Keyword(s):  
Fatty Acids ◽  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Trypanosoma Cruzi ◽  
Acid Composition ◽  
Bovine Serum ◽  
Culture Medium ◽  
Culture Media ◽  
Fetal Bovine Serum ◽  
Fetal Bovine

Fetal bovine serum (FBS) is a necessary constituent of the culture media employed to foster the growth of Trypanosoma cruzi epimastigote forms. In different laboratories, the serum is used at final concentrations of 5 or 10%. We have normally supplemented the complex medium with 10% FBS. Under this condition we have described the fatty acid composition of the total lipids and of the phosphoinositide fractions. Additionally, we have reported the increase of polyphosphoinositides and phosphatidic acid after cholinergic stimulation. Since further attempts to reproduce these results with 5% FBS in the culture medium were not successful, the effect of the FBS concentration on the fatty acid composition of phospholipids from the T. cruzi epimastigote forms was thoroughly examined. This work showed that when the FBS concentration supplementing the culture medium was reduced from 10 to 5%, the fatty acid composition of the phosphoinositides was altered while the other major phospholipids were not significantly affected. The most relevant result was the decrease in the content of linoleic acid (18:2) and the increase of palmitoleic acid (16:1) in phosphatidylinositol 4,5-bisphosphate. Phosphatidylinositol (PI) and phosphatidylinositol phosphate also exhibited similar changes in the same fatty acids. The C2fatty acid composition of the phosphoinositides, under the same conditions, is also reported here for the first time.Key words: Trypanosoma cruzi, fatty acids, phosphoinositides, fetal bovine serum, phospholipids.

High-level expression of recombinant immunoreactive thyroid peroxidase in the High Five insect cell line

1996 ◽  
Vol 17 (2) ◽  
pp. 165-174 ◽  
Author(s):  
F Grennan Jones ◽  
A Wolstenholme ◽  
S Fowler ◽  
S Smith ◽  
K Ziemnicka ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Insect Cell ◽  
Culture Medium ◽  
Aminolevulinic Acid ◽  
Expression System ◽  
Insect Cell Line ◽  
Thyroid Peroxidase ◽  
High Five Cells ◽  
Insect Cell Lines

ABSTRACT Expression of a major thyroid autoantigen, thyroid peroxidase (TPO) was studied using the baculovirus-insect cell expression system. Human TPO cDNA modified so as to code for the extracellular fragment of the protein was placed under the control of the strong polyhedrin promoter in baculovirus transfer vector pBlueBacIII and cotransfected with linearized AcMNPV viral DNA. Expression in two insect cell lines Spodoptera frugiperda (Sf9) and Tricoplusia ni (High Five) was investigated and levels of recombinant TPO (rTPO) monitored by RIA and SDS-PAGE followed by Western blotting. Both insect cell lines expressed rTPO, but higher levels (30 mg/l culture medium) were obtained with High Five cells. Culture medium rTPO was purified and its glycosylation and immunoreactivity analysed. Lectin-affinity blotting and treatment with glycosidases indicated that both high mannose and complex-type sugar residues were associated with the recombinant protein. Studies with an ELISA based on biotin-labelled rTPO and an immunoprecipitation assay based on 125I-labelled rTPO indicated that the rTPO and native TPO showed similar reactivity to TPO autoantibodies (r=0·96, P<0·001, n=50 and r=0·99, P<0·001, n=80 respectively). In addition, rTPO expressed in High Five cells showed enzyme activity comparable with that of native TPO when the heme biosynthesis precursor δ-aminolevulinic acid was included in the culture medium. Overall, our studies indicate that the High Five insect cell line provides a useful system for the expression of relatively high levels of rTPO which should be suitable for structural analysis of TPO and TPO—TPO autoantibody complexes.

scholarly journals Platelet-Rich Fibrin Extract: A Promising Fetal Bovine Serum Alternative in Explant Cultures of Human Periosteal Sheets for Regenerative Therapy

2019 ◽  
Vol 20 (5) ◽  
pp. 1053 ◽  
Author(s):  
Tomoyuki Kawase ◽  
Masaki Nagata ◽  
Kazuhiro Okuda ◽  
Takashi Ushiki ◽  
Yoko Fujimoto ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Antigen Expression ◽  
Explant Culture ◽  
The Novel ◽  
Regenerative Therapy ◽  
Platelet Rich Fibrin ◽  
Alp Activity ◽  
Fetal Bovine

In 2004, we developed autologous periosteal sheets for the treatment of periodontal bone defects. This regenerative therapy has successfully regenerated periodontal bone and augmented alveolar ridge for implant placement. However, the necessity for 6-week culture is a limitation. Here, we examined the applicability of a human platelet-rich fibrin extract (PRFext) as an alternative to fetal bovine serum (FBS) for the explant culture of periosteal sheets in a novel culture medium (MSC-PCM) originally developed for maintaining mesenchymal stem cells. Small periosteum tissue segments were expanded in MSC-PCM + 2% PRFext for 4 weeks, and the resulting periosteal sheets were compared with those prepared by the conventional method using Medium199 + 10% FBS for their growth rate, cell multilayer formation, alkaline phosphatase (ALP) activity, and surface antigen expression (CD73, CD90, and CD105). Periosteal sheets grew faster in the novel culture medium than in the conventional medium. However, assessment of cell shape and ALP activity revealed that the periosteal cells growing in the novel medium were relatively immature. These findings suggest that the novel culture medium featuring PRFext offers advantages by shortening the culture period and excluding possible risks associated with xeno-factors without negatively altering the activity of periosteal sheets.

117 Kit ligand enhances growth and nuclear maturation of buffalo oocytes in vitro

2019 ◽  
Vol 31 (1) ◽  
pp. 184
Author(s):  
M. N. Islam ◽  
M. H. Alam ◽  
A. Khatun ◽  
M. A. Hashem ◽  
M. Moniruzzaman
Keyword(s):  
Serum Sodium ◽  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Survival Rates ◽  
Sodium Pyruvate ◽  
Antral Follicles ◽  
Kit Ligand ◽  
Fetal Bovine

This study aimed to investigate the effect of Kit ligand (KL), a growth factor that regulates folliculogenesis in mammalian ovaries, on growth of buffalo oocytes in early antral follicles in vitro. Cumulus-oocyte complexes were dissected from early antral follicles (1mm) of slaughtered buffaloes and cultured in Dulbecco’s minimum essential medium supplemented with fetal bovine serum, sodium pyruvate, gentamycin, hypoxanthine, dexamethasone, cysteine, polyvinylpyrolidione, l-ascorbic acid, oestradiol-17β, and androstenedione in a 96-well culture plate at 38.5°C under an atmosphere of 5% CO2 in air for 6 days. The culture medium was supplemented with 0, 50, and 100 ng/mL KL (recombinant human SCF, Cat. No. H8416, R&amp;D Systems, Minneapolis, MN, USA). Sixty oocytes were cultured in each group with 6 replications. In vitro-grown oocytes were cultured for maturation in tissue culture medium-199 supplemented with 5% fetal bovine serum, sodium pyruvate, gentamycin, and 100 ng/mL FSH at 38.5°C for 24h under an atmosphere of 5% CO2 in air. The oocytes were then stained with aceto-orcein and examined under a differential interference contrast microscope. Data were analysed using SAS/STAT version 9.1.3 for Windows (SAS Institute Inc., Cary, NC, USA) by one-way ANOVA and means compared with Tukey’s HSD test. The mean diameter of oocytes measured at the time of seeding on the culture substrate was 100.6±0.4μm (n=180). After 6 days of culture, the diameters of oocytes increased to 110.8±0.5, 114.0±0.5, and 115.0±0.6µm in 0, 50, and 100 ng/mL KL-treated groups, respectively. The survival rates were 60.0±6, 81.2±1.2, and 92.0±4.9% in 0, 50, and 100 ng/mL KL-supplemented oocytes at Day 6. Moreover, KL pretreatment enhanced maturation of buffalo oocytes dose dependently. A small proportion of oocytes (8.4%) treated with 50 ng/mL KL reached the MII stage. This number increased to 25% when oocytes were treated with 100 ng/mL KL. These results show that KL enhances growth, viability, and meiotic progression of buffalo oocytes in vitro.

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