Horse serum

Manufactured by Cambrex

Sourced in United States

Horse serum is a laboratory product derived from the blood of horses. It is used as a supplementary component in cell culture media to support the growth and maintenance of various cell types in vitro.

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Lab products found in correlation

9 protocols using horse serum

Long-term Bone Marrow Culture Protocol

Cited 3 times

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Long-term bone marrow cultures (LTBMCs) were established according to published methods (27 (link)) from the femur and tibia marrow of 129/Sv Fanca^+/+ and 129/Sv Fanca^−/− mice at one week or one year after 7.5 Gy TBI with or without continuous 400 μM MMS350 in drinking water, as described elsewhere (18 (link)). Briefly, the contents of a femur and tibia were flushed into McCoy’s 5A medium (Thermo Fischer Scientific™, Waltham, MA) supplemented with 25% horse serum (Cambrex, Rockland, ME) and 10⁻⁵M hydrocortisone sodium hemisuccinate (Thermo Fisher Scientific). For each group, 2 mice were used to establish LTBMC and two flasks were created per mouse. Cultures were incubated at 33°C in 7% CO₂. Media was changed weekly. After 4 weeks, the horse serum was replaced with 25% fetal bovine serum (FBS; Gemini Bio-Products, West Sacramento, CA) (25 (link), 27 (link)). The cultures were observed weekly for hematopoietic cell production and adherent cell layer confluence. Nonadherent cell production data are expressed as mean ± standard error (SEM) of weekly nonadherent cell number and cumulative nonadherent cell production. Confluence data are expressed as mean ± SEM of percentage adherent cell layer confluence.

Sivananthan A., Shields D., Fisher R., Hou W., Zhang X., Franicola D., Epperly M.W., Wipf P, & Greenberger J.S. (2018). Continuous One Year Oral Administration of the Radiation Mitigator, MMS350, after Total-Body Irradiation, Restores Bone Marrow Stromal Cell Proliferative Capacity and Reduces Senescence in Fanconi Anemia (Fanca−/−) Mice. Radiation research, 191(2), 139-153.

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Hematopoietic Potential of Fancd2 Deficient Mice

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Long-term bone marrow cultures (LTBMC) were established from the femur and tibia marrow of Fancd2^+/+, Fancd2^+/− and Fancd2^−/− mice as described previously (11 (link)–13 (link)). The contents of a femur and tibia (N = 6/genotype) were flushed into McCoy’s 5A medium (Gibco, Gaithersburg, MD) supplemented with 25% horse serum (Cambrex, Rockland, ME) and 10⁻⁵M hydrocortisone sodium hemisuccinate. Cultures were incubated at 33°C in 7% CO₂. After 4 weeks, the horse serum was replaced with 25% FBS (Gibco, Gaithersburg, MD) (14 (link)). The cultures were observed weekly for hematopoietic cell production and cobblestone island formation. Cobblestone islands of greater than or equal to 50 cells were scored weekly in each flask 12 (link)–14 (link)). A two-sided two-sample t test was used to compare the number of cobblestone islands between Fancd2^+/+, Fancd2^+/− and Fancd2^−/− cultures each week. P values less than 0.05 were regarded as significant.

Berhane H., Epperly M.W., Goff J., Kalash R., Cao S., Franicola D., Zhang X., Shields D., Houghton F., Wang H., Wipf P., Parmar K, & Greenberger J.S. (2014). Radiologic Differences between Bone Marrow Stromal and Hematopoietic Progenitor Cell Lines from Fanconi Anemia (Fancd2−/−) Mice. Radiation research, 181(1), 76-89.

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Murine Long-Term Bone Marrow Cultures

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C57BL/6/NHsd and Tlr4^−/− mice were obtained from the Jackson Laboratories (B6.B10ScN-Tlr4^lps-del/JthJ, stock number 007227; Jackson Laboratories, Bar Harbor, ME, USA). LTBMCs were established from the femur and tibia marrow of mice as described elsewhere (4 (link)–8 (link)). The contents of a femur and tibia (N=6/genotype) were flushed into McCoy’s 5A medium (Gibco, Gaithersburg, MD, USA) supplemented with 25% horse serum (Cambrex, Rockland, ME, USA), and 10^–5 M hydrocortisone sodium hemisuccinate. Cultures were incubated at 33°C in 7% CO₂. After four weeks, the horse serum was replaced with 25% fetal bovine serum (FBS) (Thermo Fisher Scientific, Pittsburgh, PA, USA) (6 (link)). Cultures were observed weekly for confluence of the adherent layer of the flasks, hematopoietic cell production, and cobblestone island formation. Cobblestone islands of 50 or more cells were scored weekly in each flask (6 (link), 7 (link)). A two-sided two-sample t-test was used to compare the number of cobblestone islands between Tlr4^−/− and Tlr4^+/+ cultures each week. p-Values less than 0.05 were regarded as significant.

RHIEU B.H., EPPERLY M.W., CAO S., GOFF J., SHIELDS D., FRANICOLA D., WANG H, & GREENBERGER J.S. (2014). Improved Longevity of Hematopoiesis in Long-term Bone Marrow Cultures and Reduced Irradiation-induced Pulmonary Fibrosis in Toll-like Receptor-4 Deletion Recombinant-Negative Mice. In vivo (Athens, Greece), 28(4), 441-448.

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Culturing and Cryopreserving Cell Lines

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HeLa, NIH-3T3 and HEK-293 T cells (cryopreservation in our lab) were cultured in Dulbecco’s MEM modified medium (DMEM) with 10 % fetal bovine serum (FBS) at 37 °C and 5 % CO₂ incubator. Before being used as feeder cells, NIH-3T3 cells were exposed to 50 Gy of ⁶⁰Co radioactive source. Breast cancer cell line MCF-7 and normal mammary epithelial cell line MCF-10A were obtained from ATCC (Manassas, VA). MCF-7 cells were cultured in minimum essential medium (Eagle), supplemented with 10 % fetal bovine serum. MCF-10A cells were cultured in DMEM/F-12 (Hyclone, USA) supplemented with 10 % horse serum (Cambrex, USA), 10 μg/mL insulin, 2 μg/mL hydrocortisone, 0.01 μg/mL cholera toxin (Sigma-Aldrich, USA) and 0.02 μg/mL epidermal growth factor (EGF) (Sigma-Aldrich, USA).

Feng Z.M., Qiu J., Chen X.W., Liao R.X., Liao X.Y., Zhang L.P., Chen X., Li Y., Chen Z.T, & Sun J.G. (2015). Essential role of miR-200c in regulating self-renewal of breast cancer stem cells and their counterparts of mammary epithelium. BMC Cancer, 15, 645.

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Long-term Murine Bone Marrow Cultures

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Long-term bone marrow cultures (LTBMC) were established from the femur and tibia marrow of FVB/N background mice of genotypes, Fancd2^+/+, Fancd2^+/− and Fancd2^−/− as described by Berhane et al. (16 (link)). Briefly, the contents of a femur and tibia (N = 6/genotype) were flushed into McCoy's 5A medium (Gibco, Gaithersburg, MD) supplemented with 25% horse serum (Cambrex, Rockland, ME) and 10⁻⁵M hydrocortisone sodium hemisuccinate. Cultures were incubated at 33°C in 7% CO₂. After 4 weeks, the horse serum was replaced with 25% FBS (Gibco) (16 (link)). The cultures were observed weekly for hematopoietic cell production and cobblestone island formation. Cobblestone islands, indicative of stromal cell adherent hematopoietic islands containing primitive stem cells greater than or equal to 50 cells, were scored weekly in each flask.

Berhane H., Shinde A., Kalash R., Xu K., Epperly M.W., Goff J., Franicola D., Zhang X., Dixon T., Shields D., Wang H., Wipf P., Li S., Gao X, & Greenberger J.S. (2014). Amelioration of Radiation-Induced Oral Cavity Mucositis and Distant Bone Marrow Suppression in Fanconi Anemia Fancd2−/− (FVB/N) Mice by Intraoral GS-Nitroxide JP4-039. Radiation research, 182(1), 35-49.

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Establishment of Long-term Bone Marrow Cultures

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Long-term bone marrow cultures were established from the femurs and tibia marrow of FANCD2⁺/⁺ and FANCD2^−/−mice as described previously [15 (link),16 (link)]. Briefly, the contents of femurs and tibias were flushed into McCoy’s 5A medium (Gibco, Gaithersburg, MD, USA) supplemented with 25% horse serum (Cambrex, Rockland, ME, USA) and hydrocortisone sodium hemisuccinate. Cultures were incubated at 33°C in 7% CO2. After four weeks, the horse serum was replaced with 25% fetal bovine serum (FBS) (Gibco, Gaithersburg, MD, USA). Finally, these cells were cultured in Dulbecco’s Modified Eagle’s Medium (Invitrogen, Grand Island, NY, USA) supplemented with 10% FBS and 100U/mL penicillin (Invitrogen, Grand Island, NY, USA) in a humidified incubator with 5% CO₂ and 95% air.

Song X., Xie Y., Kang R., Hou W., Sun X., Epperly M.W., Greenberger J.S, & Tang D. (2016). FANCD2 protects against bone marrow injury from ferroptosis. Biochemical and biophysical research communications, 480(3), 443-449.

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Murine Long-Term Bone Marrow Cultures

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LTBMCs were established from the femur and tibia marrow of C57BL/6NTac mice as described elsewhere (1 (link), 2 ). The contents of a femur and tibia (n=6/genotype) were flushed into McCoy’s 5A medium (Gibco, Gaithersburg, MD, USA) supplemented with 25% horse serum (Cambrex, Rockland, ME, USA), and 10⁻⁵ M hydrocortisone sodium hemisuccinate. Cultures were incubated at 33°C in 7% CO₂. After four weeks, the horse serum was replaced with 25% fetal bovine serum (FBS) (Gibco) (1 (link), 2 ). The cultures were examined weekly for hematopoietic cell production and cobblestone island formation. Cobblestone islands of 50 cells or more were scored weekly in each flask (1 (link), 2 ). A two-sided two-sample t-test was used to compare the number of cobblestone islands between cultures each week. p-Values less than 0.05 were regarded as significant. JP4-039 was added weekly to some cultures at 10 μM as described elsewhere (11 (link)). As a vehicle control 1 μl/ml DMSO was added weekly to the cultures.

SHINDE A., EPPERLY M.W., CAO S., HOLT D., GOFF J., SHIELDS D., FRANICOLA D., WIPF P., WANG H, & GREENBERGER J.S. (2014). Improved Hematopoiesis in GS-Nitroxide (JP4-039)-treated Mouse Long-term Bone Marrow Cultures and Radioresistance of Derived Bone Marrow Stromal Cell Lines. In vivo (Athens, Greece), 28(5), 699-708.

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PC12 Cell Culture and Treatments

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PC12-G rat pheochromocytoma cells (Rausch et al. 1988 (link)) were grown in Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen, Carlsbad, CA, USA) supplemented with 7% horse serum (Cambrex, Emerainville, France), 7% heat-inactivated fetal bovine serum (Sigma), 2.5% HEPES buffer (Invitrogen), 1% glutamine (Invitrogen), 100 units/mL penicillin and 100 µg/mL streptomycin antibiotic (Invitrogen). Two days before the experiments, PC12 cells were plated on 10 cm² petri dishes or 12-well plates pre-coated with poly-L-lysine (Sigma). Cells were then cultured at 37°C in a 10% CO₂ atmosphere. Inhibitors were added 30 min prior treatment with control medium, PACAP (10⁻⁷ M) or NGF (100 ng/ml).

Seaborn T., Ravni A., Au R., Chow B.K., Fournier A., Wurtz O., Vaudry H., Eiden L.E, & Vaudry D. (2014). Induction of Serpinb1a by PACAP or NGF is required for PC12 cells survival after serum withdrawal. Journal of neurochemistry, 131(1), 21-32.

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Fancd2-Deficient Hematopoietic Stem Cell Cultures

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Long-term bone marrow cultures (LTBMC) were established from the femur and tibia marrow of (129/Sv background) Fancd2^+/+, Fancd2^+/−, and (FVB/n background) K14E7Fancd2^−/− mice according to published methods [9 (link)]. Briefly, the contents of a femur and tibia were flushed into McCoy's 5A medium (Gibco, Gaithersburg, MD) supplemented with 25% horse serum (Cambrex, Rockland, ME), and 10⁻⁵ M hydrocortisone sodium hemisuccinate. Cultures were incubated at 33°C in 7% CO₂. After 4 weeks, the horse serum was replaced with 25% FBS (Gibco, Gaithersburg, MD) [9 (link)]. The cultures were observed weekly for hematopoietic cell production and cobblestone island formation. Cobblestone islands of greater than or equal to 50 cells were scored weekly in each flask [9 (link)]. A two-sided two-sample t-test was used to compare the number of cobblestone islands between each genotype cultures each week. P-values less than 0.05 were regarded as significant.

Zhang X., Hou W., Epperly M.W., Rigatti L., Wang H., Franicola D., Sivanathan A, & Greenberger J.S. (2016). Evolution of malignant plasmacytoma cell lines from K14E7 Fancd2−/− mouse long-term bone marrow cultures. Oncotarget, 7(42), 68449-68472.

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