Sperm samples extracted from 96 mice were divided into 2 parts; the first aliquot was used for lectin histochemistry assay and the second one for enzyme assay. Each part was aliquoted into 2 subgroups. The control group was incubated with Ham's F10, and the other group was exposed to Ham's F10 (Sigma, USA) containing 75 µM of Kisspeptin (Sigma, USA) for 15 min. Then, sperm motility was checked.
Ham s f 10
Manufactured by Merck Group
Sourced in United States
Ham's F-10 is a cell culture medium used for the in vitro cultivation of a variety of cell types. It is a complex mixture of amino acids, vitamins, salts, and other nutrients required for cell growth and maintenance.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Agarose gel, by Thermo Fisher Scientific (1 mentions)
Fibroblast growth factor (fgf), by Promega (1 mentions)
Fungizone, by Thermo Fisher Scientific (1 mentions)
Basic fgf, by R&D Systems (1 mentions)
Macs separation, by Miltenyi Biotec (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
C57bl 6, by Charles River Laboratories (1 mentions)
α mem medium, by Merck Group (1 mentions)
Minimum essential medium (mem), by Merck Group (1 mentions)
Smz 2t, by Nikon (1 mentions)
Human chorionic gonadotropin (hcg), by Merck Group (1 mentions)
A549 lung cancer cells, by American Type Culture Collection (1 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Dispase, by Thermo Fisher Scientific (1 mentions)
Ham s f10, by Thermo Fisher Scientific (1 mentions)
Macs satellite cell isolation kit, by Miltenyi Biotec (1 mentions)
19 protocols using ham s f 10
1
Kisspeptin Effect on Sperm Motility
2
Effect of T. terrestris Extract on Spermatozoa
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The plant powder was thoroughly mixed with distilled water using a stirrer for 24 hours. Then, the mixture was filtered and centrifuged for 15 minutes at 5000 × g. The supernatant was collected and the solvent was evaporated under reduced pressure at 37°C. In the experimental setup using human spermatozoa, the stock solution was mixed in Ham’s-F10 (Sigma), resulting in final concentrations of 20, 40, and 50 µg/mL. Ham’s-F10 without T terrestris extract served as a control agent.
3
Sperm Motility Evaluation with Inhibitors
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Motile spermatozoa were selected from 1 ml of liquefied semen by centrifugation (600 g for 10 min) through a discontinuous Sydney IVF Sperm Gradient (Cook Medical, Bloomington, IN, USA) (0.5 ml of 80% and 0.5 ml of 40%). The supernatant was discarded and the pellet was resuspended with 1 ml of Ham's F-10 (Sigma Chemical), at pH 7.8. After another centrifugation (600 g for 5 min), the supernatant was discarded and the pellet was resuspended in 0.5 ml Ham's F-10. The sperm concentration ranged from 3 to 6 millions/ml. The sperm concentration and motility were determined by Computer Assisted Semen Analysis (CASA; Hamilton Thorne, Beverly, MA, USA). The experiments were carried out by incubating 100 ml of motile spermatozoa with 100 ml of Ham's F-10 at 37 8C, and in agreement with the experimental protocol, in the presence and absence of ouabain, amiloride, nigericin, eosin and KB-R7943 (Sigma Chemical). The sperm motility (as percentage of forward progressives from the total sperm cells analysed), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), beat cross frequency (BCF) and amplitude of lateral head displacement (ALH) were determined.
4
H9N2 Virus Plaque Assay in CEFs
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CEFs from two G2 hemizygous transgenic progeny (D-280 and E-185) and non-transgenic embryos were prepared and cultured in M199 (Invitrogen, Germany) and Ham’s F-10 (Sigma, St Louis, MO) mixed media containing 10% FBS (Hyclone, Logan, UT, USA), 0.65% sodium bicarbonate, and 1% penicillin/streptomycin. The CEFs were prepared 24 h prior to H9N2 infection at 107.5 EID50/200 µl, and the infections were performed in triplicate. After 48 h, confluent MDCK monolayers that had been propagated in 6-well plates were infected with an undiluted supernatant and a 10-fold diluted supernatant from the H9N2 virus-infected CEFs. After infection, the cells were washed with PBS and overlaid with a 0.7% agarose gel (Invitrogen) containing 1 /ml tosylsulfonyl-phenylalanyl-chloromethyl ketone (TPCK)-treated trypsin (Sigma). After 48 h, the cells were fixed in 10% PBS-buffered formalin and plaques were visualized using crystal violet staining.
5
Primary Mouse Skeletal Muscle Cell Isolation
6
Isolation and Culture of Murine Myoblasts
7
Motile Sperm Isolation via Swim-Up
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One milliliter of each sample of liquefied semen was mixed with 2 mL of Ham’s F10 (Sigma, USA), supplemented with 20% human serum albumin (HSA) (Kedrion Biopharma, Italy),
and centrifuged at 300 g for 10 min. The supernatant was decanted, and 1 mL of Ham’s F10 was added. The tubes were gently positioned at a 45-degree angle for one
hour at 37 °C and 5% CO2 for swim-up separation of motile spermatozoa. This supernatant was carefully aspirated, and its volume was adjusted to 1 mL using Ham’s F10.
and centrifuged at 300 g for 10 min. The supernatant was decanted, and 1 mL of Ham’s F10 was added. The tubes were gently positioned at a 45-degree angle for one
hour at 37 °C and 5% CO2 for swim-up separation of motile spermatozoa. This supernatant was carefully aspirated, and its volume was adjusted to 1 mL using Ham’s F10.
8
Melatonin and Luzindole Effects on Mouse Embryo Development
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Female mice were superovulated by an intra-peritoneal (IP) injection of 7.5 IU pregnant mare serum gonadotropin (PMSG, G 4877, Sigma-Aldrich, USA), followed by injection of 7.5 IU human chorionic gonadotropin (HCG, Karma, Germany) 48 hr later (10 (link)).
To ensure the reliability of gestation time, female mice were paired overnight with males of proven fertility (ratio = 1:1) and females with a vaginal plug were selected for the study.
At 42-48 hr after the HCG injection, the embryos were flushed into Ham’s F10 (Sigma, USA) medium under a stereomicroscope (Nikon SMZ-2T, Japan) (11 (link)).
After three washes, embryos with normal develop-mental morphologies were randomly divided into 3 groups including:
To ensure the reliability of gestation time, female mice were paired overnight with males of proven fertility (ratio = 1:1) and females with a vaginal plug were selected for the study.
At 42-48 hr after the HCG injection, the embryos were flushed into Ham’s F10 (Sigma, USA) medium under a stereomicroscope (Nikon SMZ-2T, Japan) (11 (link)).
After three washes, embryos with normal develop-mental morphologies were randomly divided into 3 groups including:
α-MEM medium (Sigma, USA) containing 10% fetal bovine serum (FBS) and 10-9 M melatonin as the melatonin-treated group.
α-MEM medium with serum and without melatonin as the simple media group.
α-MEM medium containing serum and 10-9 M luzindole as the Luzindole-treated group.
9
Sperm Isolation and Capacitation Protocol
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The male mice (8-12 weeks old) were sacrificed by cervical dislocation, and their cauda epididymis was excised and chopped. The sperms were capacitated in a medium (HAM’s F10 [Sigma]+4 mg/mL of bovine serum albumin [Sigma-Aldrich]) at 37°C in 5% CO2 between 45 and 60 minutes.
10
Culturing A549 Lung Cancer Cells
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A549 lung cancer cells were purchased from the American Type Culture Collection (ATCC, Manassas, USA) and maintained in DMEM/F12 medium supplemented with 10% FBS and 1% antibiotic/antimycotic mixture. Fibroblasts were obtained from skin biopsies, as described elsewhere [15 (link)], and were cultured in HAM'S F-10 (Sigma-Aldrich, St. Louis, USA) supplemented with 10% FBS, 1 mmol/l glutamine, 100 U/ml penicillin, and 100 μg/ml streptomycin at 37°C until confluence. A549 cells and fibroblasts were cultured with serum-free medium for 24 h and then the quiescent medium was changed with DMEM supplemented with 1% Nutridoma mixture (Roche) for 48h before protein extraction.
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