Ham s f 10 media

Manufactured by Merck Group

Sourced in United States, United Kingdom

Ham's F-10 media is a cell culture medium used to support the growth and maintenance of various cell types. It is a complex formulation that provides essential nutrients, vitamins, and other components necessary for cell proliferation and viability.

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Ham’s F-10 (19 citations) F-10 Ham (4 citations)

7 protocols using ham s f 10 media

Isolation and Culture of Porcine Stem Cells and Islets

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NPSCs were isolated as described previously^{11 (link),12 (link)}. Briefly, pig testes were decapsulated and chopped into 1- to 2-mm fragments in Hank’s balanced salt solution (HBSS; Sigma-Aldrich, St. Louis, MO, USA). To remove contaminating cells, tissue was digested with 2 mg/ml of sterile type V collagenase (Sigma-Aldrich), followed by trypsin 0.25% (v/v) (Roche, Madison, WI, USA) and 800 U/ml of DNAse I (Roche). Cells were cultured on non-tissue culture-treated petri dishes in supplemented Ham’s F10 media (Thermo Fisher Scientific, Waltham, MA, USA) and 10% (v/v) heat-inactivated neonatal pig serum (NPS; obtained from pigs used for islet isolation) for 48 h at 37°C to allow formation of NPSC aggregates prior to transplantation.
For isolation of NPIs, pancreases were collected from neonatal pigs and chopped using into 1- to 2-mm fragments in HBSS^{13 (link)}. Tissue was digested with 2.5 mg/ml of sterile type XI collagenase (Sigma-Aldrich) and cultured for 7–9 days on non-tissue culture-treated petri dishes in supplemented Ham’s F10 media at 37°C to purify NPIs from acinar cells^{13 (link)}.

Wright K., Dziuk R., Mital P., Kaur G, & Dufour J.M. (2016). Xenotransplanted Pig Sertoli Cells Inhibit Both the Alternative and Classical Pathways of Complement-Mediated Cell Lysis While Pig Islets Are Killed. Cell transplantation, 25(11), 2027-2040.

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Porcine Pancreatic Islet Isolation

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Neonatal porcine pancreases were procured from 3-day-old piglets (XP Bio, Anseong, Korea). The procedure for NPCC isolation was previously described by Yoon et al. [15 (link)]. Briefly, piglets were anesthetized with a mixture of rompun and ketamine (1:5 ratio). Pancreas was surgically removed, followed by cold ischemia in M199 (Thermo Fisher Scientific, Waltham, MA, USA). The pancreas was minced into less than 1 mm³ for 15 minutes, followed by enzymatic digestion with 1.5 mg/mL Collagenase P (Roche, Basel, Switzerland), dissolved in M199 for 16 minutes in a shaking water bath (37°C, 160 rpm). Digested tissues were filtered through a 500 µm cell strainer and washed using HBSS (0.25% bovine serum albumin, Sigma-Aldrich, St. Louis, MO, USA) three times. After filtration, pancreatic tissues were centrifuged for 2 minutes at 1,000 rpm. The NPCCs were then cultured in Ham’s F-10 media (Sigma-Aldrich) containing HEPES, D-glucose, L-glutamine, nicotinamide, CaCl₂, IBMX, 1× anti/anti, 10% fetal bovine serum (Thermo Fisher Scientific) at 37°C, 5% CO₂ for 5 days. Full media change was performed 24 hours later, and half the media was changed on day 3.

Cheon G.J., Park H.S., Lee E.Y., Kim M.J., You Y.H., Rhee M., Kim J.W, & Yoon K.H. (2022). Differentiation of Microencapsulated Neonatal Porcine Pancreatic Cell Clusters in Vitro Improves Transplant Efficacy in Type 1 Diabetes Mellitus Mice. Diabetes & Metabolism Journal, 46(5), 677-688.

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Isolation and Differentiation of Primary Myoblasts

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Primary myoblast cultures were isolated as previously described [36 (link)]. Briefly, hind leg muscles from 6-week-old mice were minced and digested in collagenase/dispase. Cells were collected and grown in Ham’s F-10 media (Sigma-Aldrich, St Louis, MO, USA). Medium was supplemented with 10 ng/mL bFGF. When cultures reached 70%, density medium was switched to 2% horse serum to induce differentiation. Cells were fixed 4 days after medium changed and stained using MF20 or an anti-Myogenin antibody. Fusion index was calculated as previously described [36 (link)].

Pryce B.R., Al-Zahrani K.N., Dufresne S., Belkina N., Labrèche C., Patino-Lopez G., Frenette J., Shaw S, & Sabourin L.A. (2017). Deletion of the Ste20-like kinase SLK in skeletal muscle results in a progressive myopathy and muscle weakness. Skeletal Muscle, 7, 3.

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Cell Culture of BRCA2-Deficient Cells

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Chinese hamster cells V79 (BRCA2 wild type cells) and V-C8 (brca2^−/− cells) (both gifts from Dr. Kowalczykowski) were grown in Ham's F-10 media (Invitrogen) with 15% FBS (Sigma). V-C8 cells stably expressing BRCA2 were grown in Ham's F-10 media with 15% FBS and 0.5 mg/ml G418 (Sigma). BRCA2 purification cells were grown in DMEM (Corning Cellgro) with 10% Tet-free serum (Denville Scientific) and 50 μg/ml Hygromycin B (EMD). Cells were cultured at 37°C with humidity and 5% CO₂.

Le H.P., Ma X., Vaquero J., Brinkmeyer M., Guo F., Heyer W.D, & Liu J. (2020). DSS1 and ssDNA regulate oligomerization of BRCA2. Nucleic Acids Research, 48(14), 7818-7833.

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Isolation and Sorting of Satellite Cells

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Briefly, hindlimb muscles from Ythdc1 Ctrl/iKO and Pax7n-GFP mice were dissected and minced with blades, then digested with collagenase II (1100 U/ml, Worthington in Hams F-10 media [Sigma]) for 90 min at 37°C with gentle rotation at 70 rpm. The digested muscles were washed in washing medium (Hams F-10 media, 10% HIHS [Gibco], penicillin/streptomycin [1×, Gibco]) once and SCs were further released by treating muscles with collagenase II (1100 U/ml) and dispase (11 U/ml) for 40 min at 37°C. Digested tissue was passed through a 21-gauge needle 12 times and filtered through a 40 μm filter followed by spinning at 700 × g for 5 min at 4°C. Mononuclear cells were resuspended and filtered with a 40 µm cell strainer and GFP+/EYFP+ SCs were sorted out by BD FACS Aria Fusion cell sorter (BD Biosciences).

Qiao Y., Sun Q., Chen X., He L., Wang D., Su R., Xue Y., Sun H, & Wang H. (2023). Nuclear m6A reader YTHDC1 promotes muscle stem cell activation/proliferation by regulating mRNA splicing and nuclear export. eLife, 12, e82703.

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Culturing LE2 Lung Endothelial Cells

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LE2 cells (a line of mouse lung capillary endothelia from B10D2 congenic mice, cd 133+) 44 were cultured in Hams F10 media (Sigma Aldrich, UK) supplemented with 3% FBS (Sigma Aldrich, UK), 2% antibiotic mix (60% v/v 200 mM L-glutamine (Sigma Aldrich, UK), 35% v/v penicillin/streptomycin (Sigma Aldrich, UK), 5% v/v fungizone (Invitrogen, UK)), 5 mL of 7.5% sodium bicarbonate (Sigma Aldrich, UK) and 10 mL ITS (100x, Life Technologies).
Hams was chosen as it is CO2 independent and thus can be simply cultured in different temperature environments.

Laing S., Suriano R., Lamprou D.A., Smith C.A., Dalby M.J., Mabbott S., Faulds K, & Graham D. (2016). Thermoresponsive Polymer Micropatterns Fabricated by Dip-Pen Nanolithography for a Highly Controllable Substrate with Potential Cellular Applications. ACS applied materials & interfaces, 8(37).

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Epididymal Sperm Evaluation Protocol

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Sperm samples were collected from the tail of the epididymis and then transferred to the 1 ml of Ham's F-10 media (Sigma-Aldrich Product Number N6635). After incubation at 37 °C for 20 minutes, 10 µl of the sample was placed on a slide and sperm motility was observed. The sperm count was measured by counting chamber. For examination of morphology, the sperm smear was prepared for analysis, placed on a slide, air dried at room temperature, and fixed in methyl alcohol. Then, the sample was stained with Diff-Quik.

Ziaeipour S., Ahrabi B., Naserzadeh P., Aliaghaei A., Sajadi E., Abbaszadeh H.A., Amini A., Abdi S., Darabi S, & Abdollahifar M.A. (2019). Effects of Sertoli Cell Transplantation on Spermatogenesis in Azoospermic Mice. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 52(3).

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