scholarly journals In Vitro Effects of Natrum muriaticum in Kidney Cell Lines MDCK and LLC-PK1

2021 ◽  
Vol 10 (36) ◽  
pp. 172-173
Author(s):  
Rafael Harduim ◽  
Venício Veiga ◽  
Vanessa Baldez ◽  
Marcelo Einicker-Lamas ◽  
Carla Holandino
Keyword(s):  
Cell Lines ◽  
Culture Medium ◽  
Mdck Cells ◽  
Control Cell ◽  
Calf Serum ◽  
Optical Microscope ◽  
Mitochondrial Activity ◽  
Lowry Method ◽  
In Vitro Effects

Previous papers have indicated that homeopathic solutions modify the cellular and biochemical aspects of cells maintained in culture. In this study, the effects of Natrum muriaticum, a medicine used in the homeopathic clinic for the treatment of hypertension, were evaluated in kidney MDCK and LLC-PK1 cell lines. The following cellular parameters were analyzed: viability, morphology and expression of the (Na+-K+)-ATPase and the angiotensin II receptors AT1 and AT2. The cell lines were plated (5.0 x 104 cells/mL) in DMEM supplemented with 10% fetal calf serum (FCS). After 24 hours at 37°C, DMEM was re-fed with the addition of 10% (V/V) and 1% (V/V) of the following samples: Natrum muriaticum 30CH, water 30CH and non-dynamized sterile distilled water to do the MTT assay. The results obtained from these groups were compared to those obtained by incubation of the cells in culture medium free of these solutions (Control). Cell viability was assessed by a colorimetric MTT ELISA assay (490nm). The values from four independent experiments performed in quintuplicate were plotted and statistically analyzed by Sigma Plot v.11 (Jandel Scientific). The morphology of MDCK cells was evaluated by optical microscope after Giemsa’s staining. The expression of the (Na++K+)-ATPase and AT1/AT2 of LLC-PK1 cells was evaluated by Western Blot (WB) analysis. For this experimental set, 5.0x104 cells/mL were incubated in DMEM supplemented with 10% FCS and daily culture medium was replaced by a new one, containing: Natrum muriaticum 30CH and water 30CH. Additionally, cells were treated for 5, 10 and 15 days with 1% of specific solutions and the total protein was measured by the Lowry method, after cell lysis. The samples were analyzed by electrophoresis in SDS-PAGE (12% gel) and transferred to nitrocellulose membrane. This membrane was incubated with specific primary antibodies (anti-(Na++K+)-ATPase, anti-AT1 and AT2 or anti-anti-beta-actin). The detection was performed using ECL system and Hyperfilm. MTT assays showed a statistically significant reduction in cellular mitochondrial activity (p

In vitro effects of erythromycin and florfenicol on primary cell lines of Unio crassus and Cyprinus carpio

2021 ◽  
Author(s):  
Pınar Arslan ◽  
Begum Yurdakok-Dikmen ◽  
Saniye Cevher Ozeren ◽  
Ozgur Kuzukiran ◽  
Ayhan Filazi
Keyword(s):  
Cell Lines ◽  
Cyprinus Carpio ◽  
Primary Cell ◽  
Primary Cell Lines ◽  
In Vitro Effects

In vitro effects of the chemotherapy agent water‐soluble micellar paclitaxel (Paccal Vet) on canine hemangiosarcoma cell lines

2018 ◽  
Vol 17 (1) ◽  
pp. 32-41 ◽  
Author(s):  
Caroline R. Reckelhoff ◽  
Amandine Lejeune ◽  
Patrick M. Thompson ◽  
Keijiro Shiomitsu
Keyword(s):  
Cell Lines ◽  
Water Soluble ◽  
Chemotherapy Agent ◽  
In Vitro Effects

MITOCHONDRIAL ACTIVITY OF NORMAL AND CANCER CELLS IN VITRO AT DIFFERENT CONCENTRATION OF DEUTERIUM IN CULTURE MEDIUM

2020 ◽  
pp. 139-141
Author(s):  
N. Antipova ◽  
A. Zlatska ◽  
R. Vasyliev ◽  
D. Zubov ◽  
S. Novikova ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Growth Medium ◽  
Culture Medium ◽  
Mitochondrial Activity

In the present study, we showed that normal and cancer cells in vitro in a deuterated growth medium show a decrease of mitochondrial activity (MA), while in a deuterium-depleted medium an increase. The publication has been prepared with the support of the "RUDN University Program 5-100".

In vitro effects of the activity of novel platinum (II) complex in canine and human cell lines

2019 ◽  
Vol 17 (4) ◽  
pp. 497-506 ◽  
Author(s):  
Marta Henklewska ◽  
Aleksandra Pawlak ◽  
Justyna Kutkowska ◽  
Hanna Pruchnik ◽  
Andrzej Rapak ◽  
...  
Keyword(s):  
Cell Lines ◽  
Human Cell ◽  
Human Cell Lines ◽  
In Vitro Effects

scholarly journals Structural characterization, antioxidant and cytotoxic effects of iron nanoparticles synthesized using Asphodelus aestivus Brot. aqueous extract

2020 ◽  
Vol 9 (1) ◽  
pp. 153-163 ◽  
Author(s):  
Burcu Sumer Tuzun ◽  
Tugce Fafal ◽  
Pelin Tastan ◽  
Bijen Kivcak ◽  
Besra Ozmen Yelken ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Lines ◽  
Receptor Gene ◽  
Control Cell ◽  
Antioxidant Properties ◽  
Gene Expressions ◽  
Vis Spectroscopy ◽  
Mcf 7

AbstractASP was used to synthesize FeNPA. They were characterized by UV-vis spectroscopy, FT-IR, TEM, SEM, XRD and ZP. The aim of this study was to evaluate in vitro cytotoxic activity and antioxidant acitivities of FeNPA and ASP. The antioxidant properties were evaluated using DPPH, ABTS+ and H2O2 assays. FeNPA had higher antioxidant activity comparing to ASP according to DPPH (IC50: 3.48 μg/mL) and ABTS+ (60.52%) assays. Anti-cancer activities of FeNPA and ASP were investigated in breast cancer, melanoma and control cell lines. FeNPA was more cytotoxic than ASP in MCF-7, MeWo, CHL-1, and HEL 299 cells. FeNPA had shown that mitochondria induce apoptosis through stress in MDA-MB-231, and cells MeWo. ASP also induced apoptosis 2.23-fold in MCF-7 cells. Progesterone receptor gene expression showed a 10-fold increase in a hormone-dependent MCF-7 cell line in ASP, and FeNPA treatment. Expressions of BCL6, CXCL12, DNAJC15, RB1 and TPM1 in melanoma cancer cell lines were significantly increased in ASP and FeNPA administration. It had been shown that FeNPA regulates gene expressions that may be considered important in terms of prognosis in breast cancer and melanoma cell lines and it is suggested that gene expressions regulated by FeNPA are also evaluated in animal models in vivo.

135 EXPRESSION OF APOPTOSIS-RELATED GENES IN BUFFALO (BUBALUS BUBALIS) EMBRYOS PRODUCED THROUGH IN VITRO FERTILIZATION AND PARTHENOGENETIC ACTIVATION

2011 ◽  
Vol 23 (1) ◽  
pp. 172
Author(s):  
S. Saw ◽  
K. P. Singh ◽  
R. Kaushik ◽  
M. Muzaffar ◽  
M. S. Chauhan ◽  
...  
Keyword(s):  
Relative Abundance ◽  
Culture Medium ◽  
Mammalian Species ◽  
Control Cell ◽  
Bubalus Bubalis ◽  
Blastocyst Stage ◽  
Vitro Fertilization ◽  
Immature Oocytes ◽  
Tissue Size

Apoptosis, a highly conserved evolutionary mechanism that allows an organism to tightly control cell numbers, tissue size, and protect itself from dangerous cells and unfavourable environments that threaten homeostasis, is generally directed by specific genes involved in the regulation of a series of pro-apoptotic (BAX) and anti-apoptotic (BCL-XL) proteins that are expressed during early development. All mammalian species show the highest level of spontaneous apoptotic processes at the blastocyst stage. These proteins prevent apoptosis by maintaining the cell survival by interfering with the release of cytochrome-C from mitochondria. In this study, immature oocytes were obtained from buffalo slaughterhouse ovaries and were subjected to in vitro maturation (IVM) in TCM-199 + 10% FBS + 5 μg mL–1 porcine FSH for 24 h in a CO2 incubator (5% CO2, 90 to 95% relative humidity) at 38.5°C. The mature oocytes were used for IVF, and the cleaved embryos were cultured for 8 days in culture medium (CR2 medium containing 0.6% BSA and 10% FBS) for production of embryos at different stages. The parthenotes were produced with exposure of 7% ethanol, 6-dimethyl aminopurine and cultured for 8 days in culture medium. The total RNA was isolated from oocytes and embryos and transcribed using Cell-to-cDNA-II (Ambion, Austin, TX, USA), according to manufacturer protocol. The PCR cycle included heating to 94°C for 5 min, followed by 35 cycles of 94°C for 30 s, 60 (BAX) and 62°C (BCL) for 30 s, and 72°C for 45 s with a final extension at 72°C for 10 min. The amplified product of both genes were separated on agarose gel and densitometry data for band intensities were generated using AlphaDigiDocTM AD-1201 software under a WindowsTM environment and data analysed with the help of SYSTAT software. Relative abundance of BCL-XL transcripts in immature, mature oocytes and embryos produced through IVF (i.e. 2-cell, 4-cell, 8- to 16-cell, morula, and blastocyst stage) were 25.33 ± 0.90, 12.67 ± 1.20, 37.67 ± 0.90, 30.67 ± 0.30, 23.67 ± 0.90, 18.33 ± 0.90, and 27.00 ± 1.20, respectively, whereas in parthenogenesis these values were 23.67 ± 0.88, 13.67 ± 1.20, 23.67 ± 1.20, 22.34 ± 0.88, 24.34 ± 0.88, 33.67 ± 0.88, and 45.34 ± 1.20, respectively. Relative abundance of BAX transcripts by IVF were 23.0 ± 0.60, 0.33 ± 0.10, 4.00 ± 0.60, 5.00 ± 0.60, 0.37 ± 0.06, 13.0 ± 0.66, and 56.7 ± 0.90; and by parthenonenesis were 22.3 ± 0.90, 0.13 ± 0.03, 13.67 ± 0.90, 14.0 ± 0.60, 15.33 ± 0.90, 64.67 ± 2.20, and 55.0 ± 2.10, respectively. In conclusion, the expression pattern of the apoptosis-related genes revealed that the incidence of apoptosis was significantly higher in IVF and parthenogenetically produced buffalo embryos at stages such as immature oocytes, morula, and blastocyst than the early cleavage stage embryos.

232 ADDITION OF VERY LOW AMOUNTS OF SERUM (ESTRUS COW SERUM) IMPROVES IN VITRO EMBRYO PRODUCTION IN DAIRY CATTLE

2015 ◽  
Vol 27 (1) ◽  
pp. 205 ◽  
Author(s):  
E. Mullaart ◽  
F. Dotinga ◽  
C. Ponsart ◽  
H. Knijn ◽  
J. Schouten
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Calf Serum ◽  
High Serum ◽  
In Vitro Production ◽  
Embryo Production ◽  
Chi Square ◽  
Embryo Yield ◽  
Vitro Production

Improving the efficiency of the in vitro production (IVP) process is very important because it results in more embryos to be used in breeding programs or as commercial service. At CRV, a culture medium consisting of SOF with amino acids and BSA is used. In the past, richer culture media were used with 10% fetal calf serum combined with BRL cell co-culture. Although the efficiency of the IVP process of these media was good, these rather high serum concentrations were quite often related to large offspring syndrome (LOS). The switch to a culture system without serum resulted in a significant reduction in LOS but also in a reduction of embryo yield. The aim of the present study was to investigate the effect of adding low amounts of serum to the culture medium on efficiency of embryo production. Immature cumulus-oocyte complexes (COC) were recovered from ovaries 6 to 8 h upon slaughter. The COC were matured in vitro in TCM199/FCS/LH/FSH supplemented with cysteamine (0.1 mM). Subsequently, matured oocytes were fertilised with frozen-thawed gradient-separated semen and further cultured for 7 days in SOFaaBSA. The SOF medium contained either 0 (control), 0.1, 0.5, or 1.0% oestrus cow serum (ECS). Embryos development was scored at Day 7. Three replicates were performed and results were analysed by chi-square analyses. The results clearly show that adding ECS significantly improved embryo production (Table 1). Interestingly, already very low amounts (0.1%) of serum gave a significant increase in embryo percentage. In conclusion, addition of very low amounts of ECS (0.1%) is beneficial for embryo production, resulting in significantly higher embryo production (from 19 to 27%). In a subsequent field trial with OPU-derived embryos, the effect of addition of 0.1% ECS on birth weight (LOS) of the calves has to be investigated. Table 1.Percentage of blastocysts at Day 7 after culture in SOF medium with different amounts of serum

92 Culture method for long-distance transport of bovine embryos derived from IVF before blastulation using microtubes

2019 ◽  
Vol 31 (1) ◽  
pp. 172
Author(s):  
T. Yamanouchi ◽  
H. Matsuda ◽  
K. Ogata ◽  
Y. Hashiyada
Keyword(s):  
Culture Medium ◽  
Liquid Paraffin ◽  
Calf Serum ◽  
Petri Dish ◽  
Culture Method ◽  
Optimal Number ◽  
Blastocyst Development ◽  
Long Distance

In vitro-produced (IVP) embryos are more easily damaged by cryopreservation than in vivo-derived embryos. Therefore, transportation of fresh IVP embryos in a manner that can maintain viability is necessary. This study was conducted to determine the preferable culture conditions for transport of embryos at 5 days post-insemination (dpi) in 1.5-mL microtubes. Cumulus-oocyte complexes derived from an abattoir were matured and then inseminated with frozen-thawed semen. Presumptive zygotes were cultured in mCR1aa (CR1)+5% calf serum (CS) until use. In Exp. 1, embryos with 5 blastomeres at 5 dpi were randomly assigned to 1 of 3 groups: 25mM Hepes-CR1aa (H-CR1)+5% CS or 25mM Hepes-M199 (H-M199)+5% CS in air, or CR1 in 5% CO2. Embryos were cultured in microdrops overlaid with liquid paraffin in a petri dish for 48h at 38.5°C. In Exp. 2, the optimal number of embryos to culture per microtube was assessed. Presumptive zygotes were cultured in groups of 20, 40, or 80 in 1mL of CR1 covered with liquid paraffin in microtubes in an incubator at 38.5°C in 5% CO2 until 7 dpi. For Exp. 3, culture of embryos in microtubes in a portable incubator was tested. At 5 dpi, 5-cell embryos (n=17 to 36 per microtube) were statically cultured in 1mL of CR1 or H-CR1 in microtubes in a portable incubator set at 38.5°C for 48h. The CR1 was pre-equilibrated in an incubator in 5% CO2 for 24h before use. Embryos were harvested from microtubes after 48h and were then cultured in microdrops of CR1 overlaid with liquid paraffin in a petri dish in an incubator at 38.5°C in 5% CO2 until 8 dpi. In Exp. 4, embryos (n=29 to 39 five-cell embryos per microtube) were transported in a portable incubator by land for 1000km over a period of 44h using the same conditions as in Exp. 3. Control embryos were statically cultured in microdrops of CR1 in an incubator in 5% CO2. Statistical analyses were carried out by ANOVA (Exp. 1 and 2), t-test (Exp. 3), or Fisher’s exact test (Exp. 4). In Exp. 1, there was no effect (P>0.05) of culture medium on blastocyst development at 7 dpi (27.6±2.3, 25.7±7.2, and 17.3±2.9% for CR1, H-CR1, and H-M199, respectively). In Exp. 2, blastocyst development at 7 dpi was not affected (P>0.05) by the number of presumptive zygotes cultured per microtube (43.6±8.3, 42.4±4.0, and 39.9±2.9% for 20, 40, and 80 presumptive zygotes, respectively). In Exp. 3, blastocyst development at 8 dpi was not affected (P>0.05) by culture medium (60.7±7.4 and 53.1±4.4% for CR1 and H-CR1, respectively); however, the pH of CR1 changed from 7.5 to 8.1 at 48h after culture. In Exp. 4, blastocyst development at 8 dpi was not affected (P>0.05) by transport (57.1, 64.4, and 75.5% for CR1, H-CR1, and control, respectively). These results indicate that IVP embryos harvested at 5 dpi can be transported by portable incubator with no effect on embryo development to the blastocyst stage. This work was supported by grants from the Project of the Bio-oriented Technology Research Advancement Institution, NARO (the special scheme project on advanced research and the development for next-generation technology).

Über Antigenstrukturen von permanenten Zellstämmen

1960 ◽  
Vol 15 (11) ◽  
pp. 707-713 ◽  
Author(s):  
Arno Geissler ◽  
Else Knake
Keyword(s):  
Culture Medium ◽  
Complement Fixation Test ◽  
Complement Fixation ◽  
Calf Serum ◽  
Antigenic Property ◽  
Fixation Test ◽  
Established Cell ◽  
Cell Strains ◽  
Original Species

Using the complement fixation test, we found three different antigenic components in established cell strains originating from man and mouse:1. the antigenic property of the original species specifity. It resides in cell substances which, in a serological sense, are haptenes.2. an acquired antigenic property of species specifity of beef, which is a haptene too. The cells acquired this antigenic component from substances in the culture medium containing 20% of calf serum.3. an antigenic property characterized as a complete antigen which, probably, was acquired by the cells only during their in vitro-life. The nature of this third antigen is not yet entirely clear.

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