Ham s f10 nutrient mixture

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom

Ham's F10 nutrient mixture is a cell culture media formulation developed for the growth and maintenance of various cell types. It provides a balanced combination of amino acids, vitamins, salts, and other essential components to support cell proliferation and viability in in vitro cell culture applications.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions) Glutamax, by Thermo Fisher Scientific (3 mentions) Horse serum, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Merck Group (2 mentions) L glutamine, by Merck Group (2 mentions) Axiolab live cell imager, by Zeiss (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) B fgf, by Promega (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions) Trypsin edta, by Welgene (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Hygromycin b, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Lonza (1 mentions) Dispase 2, by Merck Group (1 mentions) Collagenase 1, by Thermo Fisher Scientific (1 mentions) Aphidicolin, by Merck Group (1 mentions) Penicillin, by Corning (1 mentions) Streptomycin, by Corning (1 mentions)

15 protocols using ham s f10 nutrient mixture

Isolation and Culture of Avian and Murine Myoblasts

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Chicken primary myoblasts were isolated from the chicken leg and breast muscle of day 10 embryo as previous described.¹² Primary myoblast represented the chicken primary myoblasts that have just completed serial plating. Growing myoblast represented myoblasts that were cultured in growth medium with RPMI‐1640 (Gibco, Grand Island, NY, USA), 15% foetal bovine serum (FBS) (ExCell, Shanghai, China), 10% chicken embryo extract, and 0.2% penicillin/streptomycin (Gibco) at 37°C in 5% CO₂. Differentiated myotube (DM) represented myoblasts induced to differentiation for 4 days by culturing the cells in differentiation medium (RPMI‐1640 without FBS containing 2% horse serum) when 90% confluent.
Mouse primary myoblasts were isolated and cultured as previously described.¹³ Cells were isolated from the forelimbs and hindlimbs of 3‐week‐old mice, minced and digested in a solution of dispase B and type I collagenase. Growth medium consisted of Ham's F‐10 nutrient mixture (Gibco) supplemented with 20% FBS (ExCell) and 2.5 ng/mL bFGF (Promega, Madison, WI, USA). Differentiation medium consisted of DMEM (Gibco) supplemented with 2% horse serum (Gibco). All medium contained 0.2% penicillin/streptomycin (Gibco).

Luo W., Lin Z., Chen J., Chen G., Zhang S., Liu M., Li H., He D., Liang S., Luo Q., Zhang D., Nie Q, & Zhang X. (2021). TMEM182 interacts with integrin beta 1 and regulates myoblast differentiation and muscle regeneration. Journal of Cachexia, Sarcopenia and Muscle, 12(6), 1704-1723.

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Isolation and Culture of Muscle-Derived Primary Cells

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Five times of 10⁵ muscle-derived primary cells were seeded onto 35-mm culture dishes (SPL) and cultured in Ham’s F10 nutrient mixture (Gibco, Grand Island, NY, USA ) supplemented with 15% (v/v) FBS, 2.5 ng/ml basic fibroblast growth factor (Peprotech, Rocky Hill, NJ, USA), and 1% (v/v) antibiotic-antimycotic (herein referred to as proliferation medium). After culture for 24 h, floating (non-adherent) cells were collected and centrifuged at 1,500×g for 4 min. The pellets were then re-suspended in fresh proliferation medium and incubated in 35-mm culture dishes at 37℃ in a humidified atmosphere of 5% CO₂ in air. After 72 h, adherent cells were retrieved via treatment with 0.25% trypsin-EDTA (Welgene), washed with fresh proliferation medium, and used for the following experiments. Details of the DP method are shown in Fig. S1.

Lee H., Han N.R., Kim S.J., Yun J.I, & Lee S.T. (2022). Development of a High-Yield Isolation Protocol Optimized for the Retrieval of Active Muscle Satellite Cells from Mouse Skeletal Muscle Tissue. International Journal of Stem Cells, 15(3), 283-290.

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Stable Transfection of α7 nAChR in GH4C1 Cells

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GH4C1 cells stably transfected with pCEP4/rat α7 nAChR were obtained from Siena Biotech S.p.A. (Siena, Italy). GH4C1 cells endogenously express the assembly promoting chaperone protein RIC-3 (Williams et al., 2005 (link)). Cells were cultured in poly-L-lysine (PLL)-coated standard tissue culture flasks (T-25) at 37 °C in 95% humidified air and 5% CO₂. Culture medium contained Ham’s F10 Nutrient Mixture (Gibco), 15% horse serum (Gibco), 2.5% fetal bovine serum (Gibco), 1% penicillin–streptomycin (Lonza), 1 mM GlutaMAX (Gibco) and 100 µg/ml Hygromycin B (Invitrogen). Before the experiments, at ∼80% confluency, the cells were plated onto PLL-coated Petri dishes (d = 35 mm) for use on the following day.

Pesti K., Lukacs P, & Mike A. (2019). Type I-like behavior of the type II α7 nicotinic acetylcholine receptor positive allosteric modulator A-867744. PeerJ, 7, e7542.

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Isolation and Differentiation of Mouse Myoblasts

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Mouse myoblasts were isolated as described previously with some modifications (17 (link)). Briefly, hindlimbs of 2-day-old C57BL/6J mice were removed and muscle was separated from the skin and bone with sterile forceps. The cells were dispersed with 0.2% collagenase I (Gibco) and 0.2% Dispase II (Sigma-Aldrich) for 30 min, and then filtered through a 40 μm cell strainer. To remove non-myogenic cells, cells were plated on normal culture dish and incubated for 30 min. Subsequently, floating cells were re-plated on a collagen coated culture dish. Myoblasts were initially cultured in Ham's F10 nutrient mixture (Gibco) and then the medium was switched to DMEM/Ham's F10 (1:1). Both growth media were supplemented with 20% FBS, 2.5 ng/ml bFGF and 1% penicillin/streptomycin. To induce differentiation, myoblasts were cultured in DMEM with 5% HS and penicillin/streptomycin. Medium was changed every other day.

Anan K., Hino S., Shimizu N., Sakamoto A., Nagaoka K., Takase R., Kohrogi K., Araki H., Hino Y., Usuki S., Oki S., Tanaka H., Nakamura K., Endo F, & Nakao M. (2018). LSD1 mediates metabolic reprogramming by glucocorticoids during myogenic differentiation. Nucleic Acids Research, 46(11), 5441-5454.

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Culturing Immortalized BJ Cells

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BJ cells (ATCC), and derivatives, were cultured in Ham's F10 nutrient mixture (Life Technologies, Grand Island, NY, USA) supplemented with 15% batch-tested fetal bovine serum (Atlanta Biologicals, Lawrenceville, GA, USA), 20 mm L-glutamine (Cellgro, Manassas, VA, USA), 100 U ml⁻¹ penicillin, and 100 μg ml⁻¹ streptomycin (Cellgro). BJ-hTERT cells were generated by retroviral transduction of BJ cells using the pBabe-hTERT-puro vector followed by drug selection. Cultures were passaged at 1:4 and incubated at 37 °C in atmosphere of 5% CO₂ and 2% or 21% Oxygen as indicated. Cells were labeled with 1 μg ml⁻¹ BrdU (GE Healthcare, Piscataway, NJ, USA), and aphidicolin (Sigma, St. Louis, MO, USA; 0.2 μm) was directly added to the culture medium. Cell proliferation curves were generated by counting cells using a hemocytometer and the formula PD = log2(N_final/N_initial), where N_initial is the number of cells seeded at each passage and N_final is the number of cells recovered from the dish.

Boccardi V., Razdan N., Kaplunov J., Mundra J.J., Kimura M., Aviv A, & Herbig U. (2015). Stn1 is critical for telomere maintenance and long-term viability of somatic human cells. Aging Cell, 14(3), 372-381.

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Density Gradient Sperm Purification and Cryopreservation

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Ham's F-10 nutrient mixture (Life Technologies, Paisley, UK) supplemented with 0.6% (w/v) bovine serum albumin (BSA) and 26 mM bicarbonate (NaHCO3) was used to wash all the semen samples at room temperature. Afterwards, the samples were treated by density gradient centrifugation at room temperature, with 80% (v/v) and 40% (v/v) Percoll at a speed of 400 g for 30 minutes. All sperm cell specimens were examined using a phase-contrast microscope, and no other impurities were detected. As the number of sperm cells obtained by centrifugation was inadequate, the spermatozoa of three individuals were combined into one sample. As a result, there were ten samples for each of the IAS and normozoospermic groups. Spermatozoa were washed with cold PBS three times, cryopreserved in liquid nitrogen, and stored at −80°C until further metabolite extraction was performed.

Zhao K., Zhang J., Xu Z., Xu Y., Xu A., Chen W., Miao C., Liu S., Wang Z, & Jia R. (2018). Metabolomic Profiling of Human Spermatozoa in Idiopathic Asthenozoospermia Patients Using Gas Chromatography-Mass Spectrometry. BioMed Research International, 2018, 8327506.

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Sperm Metabolomic Sample Preparation

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All semen samples were washed using Ham's F-10 nutrient mixture (Life Technologies, Paisley, UK) supplemented with 0.6% (w/v) bovine serum albumin (BSA) and 26 mM bicarbonate (NaHCO 3 ) at room temperature. Samples were then prepared by density gradient centrifugation, using 80% (v/v) and 40% Percoll (GE Healthcare, Uppsala, Sweden) for 30 min at 400 g, at room temperature. The purity of all sperm samples was checked using phase contrast microscopy and only samples with no detectable contamination with non-sperm cells after processing were used for metabolomic analyses.
Sperm cells were then washed three times with cold PBS and, depending on the metabolomics technique used, cells were frozen with liquid nitrogen and stored at À80 °C for further metabolite extraction (for gas chromatography (GC) time-of flight (TOF)/mass spectrometry (MS) analysis) or metabolites were directly extracted and lyophilized (for proton nuclear magnetic resonance -1 H-NMR)).

Paiva C., Amaral A., Rodriguez M., Canyellas N., Correig X., Ballescà J.L., Ramalho-Santos J, & Oliva R. (2015). Identification of endogenous metabolites in human sperm cells using proton nuclear magnetic resonance ((1) H-NMR) spectroscopy and gas chromatography-mass spectrometry (GC-MS). Andrology, 3(3).

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Cytokine-Induced Aggregation Assay

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At the end of the differentiation, cell aggregates were resuspended from microwell plates, transferred into 10-cm Petri dishes, and manually collected using a micropipette (200 aggregates per condition). The retrieved aggregates were washed twice with pre-warmed PBS and resuspended in 1 mL of HAM’s F-10 nutrient mixture (Thermofisher, Waltham, MA, USA) containing 3.75 g free fatty acid-free BSA (bovine serum albumin) (Roche, Basel, Switzerland), 2.5 mL GlutaMAX (Thermofisher), and 100 U/mL penicillin-streptomycin (Thermofisher), and cytokines were added (1000 U/mL IFNγ (Peprotech, London, UK) + 50 U/mL IL-1β (R&D Systems, Abingdon, UK) [16 (link)] or 2000 U/mL IFNα (Peprotech) [17 (link)]). Cells were exposed to cytokines for 24 or 48 h, as described in figure legends.

Demine S., Schiavo A.A., Marín-Cañas S., Marchetti P., Cnop M, & Eizirik D.L. (2020). Pro-inflammatory cytokines induce cell death, inflammatory responses, and endoplasmic reticulum stress in human iPSC-derived beta cells. Stem Cell Research & Therapy, 11, 7.

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Murine Skeletal Muscle Stem Cell Expansion and Differentiation

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Petri dishes were pre-coated with 0.5 mg/mL type I collagen (Corning, cat# 354236, 0.02 M dissolved in acetic acid) overnight, then washed three times with PBS. Ham’s F-10 Nutrient Mixture (Thermo Fisher Scientific, cat# 11550043) with 20% FBS, 1% penicillin/streptomycin, and 10 ng/mL FGF (R and D Systems, cat# 233-FB-025) was used as the base medium. For in vitro expansion, MuSCs were cultured in 50% volume of F-10 base medium combined with 50% volume of conditioned medium (NCM, ACM, or F-10 medium). Cells were passaged every 48 h. For differentiation induction, MuSCs were washed three times with PBS and changed into differentiation medium (DMEM containing 2% horse serum, HyClone, cat# HYCLSH30074.03HI, and 1% penicillin/streptomycin) for 48 h.

Zhou S., Zhang W., Cai G., Ding Y., Wei C., Li S., Yang Y., Qin J., Liu D., Zhang H., Shao X., Wang J., Wang H., Yang W., Wang H., Chen S., Hu P, & Sun L. (2020). Myofiber necroptosis promotes muscle stem cell proliferation via releasing Tenascin-C during regeneration. Cell Research, 30(12), 1063-1077.

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Isolation of Mouse Skeletal Muscle Progenitor Cells

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Skeletal muscle tissues were removed from the limbs of neonatal mice. The tissues were cut into small pieces and digested with the enzyme mixture (1.5 U/mL collagenase D) (Roche) (Basel, Switzerland), 2.5 mM CaCl₂ and 2 U/mL Dispase (Roche) (Basel, Switzerland) in serum-free DMEM at 37 °C for 0.5–1 h. Ten milliliters of DMEM was added to the digested mixture and triturated prior to centrifugation at 1500× g for 10 min at room temperature. The pellets were resuspended in Ham’s F-10 nutrient mixture (Thermofisher) (Waltham, MA, USA) containing 20% FBS, bFGF (R&D) (New York, USA) (5 ng/mL) and 1% penicillin/streptomycin and plated in a 10 cm culture dish for 1 h. The unattached cells were harvested and cultured in 24-well culture plates that were precoated with Matrigel (Sigma-Aldrich) (St. Louis, MO, USA).

Li J., Shi M., Liu L., Wang J., Zhu M, & Chen H. (2022). Tetrandrine Inhibits Skeletal Muscle Differentiation by Blocking Autophagic Flux. International Journal of Molecular Sciences, 23(15), 8148.

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