scholarly journals Milk of Cow (Bos taurus), Buffalo (Bubalus bubalis), and Goat (Capra hircus): a Better Alternative than Fetal Bovine Serum in Media for Primary Isolation, In Vitro Cultivation, and Maintenance of Leishmania donovani Promastigotes

2007 ◽  
Vol 45 (4) ◽  
pp. 1353-1356 ◽  
Author(s):  
M. Muniaraj ◽  
C. S. Lal ◽  
S. Kumar ◽  
P. K. Sinha ◽  
P. Das
Keyword(s):  
Bovine Serum ◽  
Leishmania Donovani ◽  
Bos Taurus ◽  
Fetal Bovine Serum ◽  
Bubalus Bubalis ◽  
Capra Hircus ◽  
In Vitro Cultivation ◽  
Primary Isolation ◽  
Fetal Bovine

Soy protein isolate: A substitute of fetal bovine serum for the in vitro cultivation of Leishmania donovani

2017 ◽  
Author(s):  
Arushdeep Sidana ◽  
Afroz Alam ◽  
Umar Farooq
Keyword(s):  
Soy Protein ◽  
Bovine Serum ◽  
Culture Medium ◽  
Leishmania Donovani ◽  
Fetal Bovine Serum ◽  
Soy Protein Isolate ◽  
Protein Isolate ◽  
In Vitro Cultivation ◽  
Fetal Bovine

Fetal Bovine Serum (FBS) is an expensive source of macronutrients which are required for proper nutrition of Leishmania parasite in the culture medium. An alternative, cost effective source of macronutrients which can replace the use of FBS in tissue culture medium is required. The potential of Soy Protein Isolate (SPI) to replace FBS in RPMI-1640 medium for the in vitro cultivation of Leishmania donovani was evaluated. Commercially available SPI powder was used in RPMI-1640 medium as a substitute of FBS to cultivate L. donovani promastigotes. The growth, multiplication and morphology of cultivated parasites was observed in conventional RPMI-1640 with 10% FBS (v/v) and RPMI-1640 containing 10% SPI (v/v) by using light microscopy, measurement of absorbance and cell counting. The growth of Leishmania promastigotes in the medium containing 10% SPI was slower in initial phase; however, the parasites were morphologically larger as compared to those in RPMI-1640 medium containing 10% FBS. Cell count in the SPI-containing RPMI-1640 medium was 2.3 × 108 cells/ml whereas it was 1.9 × 107 cells/ml in RPMI-1640 with 10% FBS. This study concludes that RPMI-1640 may be supplemented with SPI instead of FBS for the in vitro cultivation of Leishmania donovani promastigotes to decrease the culture maintenance cost in developing countries.

90 OVIDUCTAL EPITHELIAL CELLS Co-CULTURE PROMOTES GOAT (CAPRA HIRCUS) IN VITRO PARTHENOGENETIC EMBRYO DEVELOPMENT

2017 ◽  
Vol 29 (1) ◽  
pp. 152
Author(s):  
M. Mahajan ◽  
D. Nagoorvali ◽  
N. Rawat ◽  
M. S. Chauhan ◽  
R. S. Manik ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Calcium Ionophore ◽  
Mrna Abundance ◽  
Developmental Competence ◽  
Capra Hircus ◽  
Fetal Bovine

Co-culture of pre-implantation embryos with oviducal epithelial cells mimics the in vivo conditions, thus, playing a crucial role in embryo metabolism and gene expression and finally supporting embryonic developmental competence in several ways. Hence, the objective of the present study was to evaluate the effect of goat oviducal epithelial cells (GOEC) co-culture on goat parthenogenetic embryonic development, quality, and relative mRNA abundance of genes related to developmental competence and oxidative stress. The GOEC were obtained from goat oviducts by squeezing and thorough washing with TCM-199 + 10% fetal bovine serum. Goat cumulus–oocyte complexes were collected from slaughterhouse ovaries and matured in TCM-199 + 10% fetal bovine serum supplemented with 5 μg mL−1 of FSH, 10 μg mL−1 of LH, and 1 μg mL−1 of β-oestradiol for 27 h in CO2 incubator with 5% CO2 and at 38.5°C with >95% RH. In vitro matured cumulus–oocyte complexes were denuded and activated with 5 μM calcium ionophore and 2 mM 6-DMAP. Following activation, embryos were co-cultured with and without GOEC (control) in mCR2aa media. The blastocyst development rate was significantly (P < 0.05) higher (23.00 ± 1.15% v. 17.33 ± 1.45%) in the media cultured with GOEC than in control. The total cell number of blastocysts (n = 4) was also found to be significantly more (167.25 ± 17.51 v. 110.25 ± 12.02) than that of control (P < 0.05). However, the apoptotic index (3.76 ± 0.23% v. 7.97 ± 1.99%) was not significantly different in both groups. Further, RNA was isolated from both groups (20 each) of blastocysts on Day 8, and cDNA was prepared. Analysis by qPCR revealed that the relative mRNA abundance of development related genes, i.e. VEGF, BMP4, and CCNB1, showed significantly high (P < 0.05) expression, whereas the expression of CRABP1 was significantly low (P < 0.05) in GOEC co-culture than control. Oxidative stress related genes GPX-1 and SOD2 had comparable expression in both the culture systems, whereas a nonsignificant (P < 0.05) increase in expression of PRDX1 was observed in GOEC co-culture group. In conclusion, co-culture of embryos with GOEC in the simple culture media like mCR2aa helps in improving developmental competence and quality of parthenogenetic embryos. This work was supported by the NFBSFARA Project on Parthenogenetic Goat (CA-4002), New Delhi, India.

scholarly journals Effects of fetal bovine serum and estrus buffalo serum on maturation of buffalo (Bubalus bubalis) oocytes in vitro

2015 ◽  
Vol 8 (2) ◽  
pp. 143-146 ◽  
Author(s):  
Gopal Puri ◽  
S. S. Chaudhary ◽  
V. K. Singh ◽  
A. K. Sharma
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Bubalus Bubalis ◽  
Fetal Bovine

Isolation and in vitro cultivation of the aphid pathogenic fungusEntomophthora planchoniana

2001 ◽  
Vol 47 (12) ◽  
pp. 1082-1087 ◽  
Author(s):  
Florian M Freimoser ◽  
Annette B Jensen ◽  
Urs Tuor ◽  
Markus Aebi ◽  
Jørgen Eilenberg
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Large Subunit ◽  
Small Subunit ◽  
In Vitro Cultivation ◽  
Small Subunit Rdna ◽  
Fetal Bovine ◽  
Large Subunit Rdna ◽  
First Time

Entomophthora planchoniana is an important fungal pathogen of aphids. Although Entomophthora chromaphidis has been considered a synonym for E. planchoniana, the two species are now separated, and E. planchoniana is reported not to grow in vitro. In this paper, we describe for the first time the isolation and cultivation of this species. Entomophthora planchoniana was isolated from a population of Ovatus crataegarius (Homoptera, Aphididae), which was infected by E. planchoniana only. The isolates did not sporulate, but the sequence of the small subunit rDNA and the restriction fragment length polymorphism patterns of the first part of the large subunit rDNA and the ITS II region confirm that the isolates were E. planchoniana. The isolated fungus grew in a medium consisting of Grace's insect cell culture medium supplemented with lactalbumin hydrolysate, yeastolate, and 10% fetal bovine serum or in GLEN medium with 10% fetal bovine serum. Vegetative cells of E. planchoniana were long and club-shaped and did not stain with Calcofluor, thus suggesting that they were protoplasts.Key words: biological control, entomopathogen, Entomophthorales, protoplast, Zygomycetes.

Successful piglet production by IVF of oocytes matured in vitro using NCSU-37 supplemented with fetal bovine serum

2006 ◽  
Vol 65 (2) ◽  
pp. 374-386 ◽  
Author(s):  
Misae Suzuki ◽  
Koji Misumi ◽  
Manabu Ozawa ◽  
Junko Noguchi ◽  
Hiroyuki Kaneko ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Fetal Bovine

scholarly journals Stimulatory Role of Glycated Fetal Bovine Serum Along with Iron on In Vitro Production of Insulin by Differentiated Mouse Embryonic Stem Cells

2011 ◽  
Vol 57 (4) ◽  
pp. 356-361
Author(s):  
Ikuo Nishigaki ◽  
Gowri Rangasamy Gunassekaran ◽  
Panjan Nagappan Venkatesan ◽  
Mandupal Chaco Sabu ◽  
Sabu Priya ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Embryonic Stem ◽  
In Vitro Production ◽  
Fetal Bovine ◽  
Vitro Production

scholarly journals Fetal Bovine Serum-Derived Extracellular Vesicles Persist within Vesicle-Depleted Culture Media

2018 ◽  
Vol 19 (11) ◽  
pp. 3538 ◽  
Author(s):  
Brandon Lehrich ◽  
Yaxuan Liang ◽  
Pooya Khosravi ◽  
Howard Federoff ◽  
Massimo Fiandaca
Keyword(s):  
Cell Growth ◽  
Extracellular Vesicles ◽  
Bovine Serum ◽  
Culture Media ◽  
Fetal Bovine Serum ◽  
Cellular Growth ◽  
Primary Astrocyte ◽  
Fetal Bovine ◽  
Cell Growth And Viability

It is known that culture media (CM) promotes cellular growth, adhesion, and protects explanted primary brain cells from in vitro stresses. The fetal bovine serum (FBS) supplement used in most CM, however, contains significant quantities of extracellular vesicles (EVs) that confound quantitative and qualitative analyses from the EVs produced by the cultured cells. We quantitatively tested the ability of common FBS EV-depletion protocols to remove exogenous EVs from FBS-supplemented CM and evaluated the influence such methods have on primary astrocyte culture growth and viability. We assessed two methodologies utilized for FBS EV removal prior to adding to CM: (1) an 18-h ultracentrifugation (UC); and (2) a commercial EV-depleted FBS (Exo-FBS™). Our analysis demonstrated that Exo-FBS™ CM provided the largest depletion (75%) of total FBS EVs, while still providing 6.92 × 109 ± 1.39 × 108 EVs/mL. In addition, both UC and Exo-FBS™ CM resulted in poor primary astrocyte cell growth and viability in culture. The two common FBS EV-depletion methods investigated, therefore, not only contaminate in vitro primary cell-derived EV analyses, but also provide a suboptimal environment for primary astrocyte cell growth and viability. It appears likely that future CM optimization, using a serum-free alternative, might be required to advance analyses of cell-specific EVs isolated in vitro.

scholarly journals Profiling of human neural crest chemoattractant activity as replacement of fetal bovine serum for in vitro chemotaxis assays

2021 ◽  
Author(s):  
Xenia Dolde ◽  
Christiaan Karreman ◽  
Marianne Wiechers ◽  
Stefan Schildknecht ◽  
Marcel Leist
Keyword(s):  
Neural Crest ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Developmental Toxicity ◽  
Video Tracking ◽  
Large Panel ◽  
Human Proteins ◽  
Fetal Bovine ◽  
Targeted Approach

Fetal bovine serum (FBS) is the only known stimulus for migration of human neural crest cells (NCCs). Non-animal chemoattractants are desirable for the optimization of chemotaxis assays to be incorporated in a test battery for reproductive and developmental toxicity. We confirmed here in an optimized transwell assay that FBS triggers directed migration along a concentration gradient. The responsible factor was found to be a protein in the 30-100 kDa size range. In a targeted approach, we tested a large panel of serum constituents known to be chemotactic for NCCs in animal models (e.g. VEGF, PDGF, FGF, SDF-1/CXCL12, ephrins, endothelin, Wnt, BMPs). None of the corresponding human proteins showed any effect in our chemotaxis assays based on human NCCs. We then examined in a broad screening approach, whether human cells would produce any factor able to trigger NCC migration. We found that HepG2 hepatoma cells produced chemotaxis-triggering activity (CTA). Using chromatographic methods and by employing the NCC chemotaxis test as bioassay, the responsible protein was enriched by up to 5000-fold. We also explored human serum and platelets as direct source, independent of any cell culture manipulations. A CTA was enriched from platelet lysates several thousand-fold. Its temperature and protease-sensitivity suggested a protein component. The capacity of this factor to trigger chemotaxis was confirmed by single-cell video-tracking analysis of migrating NCCs. The human CTA characterized here may be employed in the future for the setup of assays testing for the disturbance of directed NCC migration by toxicants.

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