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Murine stem cell factor

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Murine stem cell factor is a recombinant protein that promotes the survival and proliferation of murine (mouse) hematopoietic stem and progenitor cells. It is used as a cell culture supplement in research applications.

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Lab products found in correlation

Murine il 3, by Thermo Fisher Scientific (6 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) Interleukin 3 (il 3), by Thermo Fisher Scientific (3 mentions) Murine interleukin 3, by Thermo Fisher Scientific (3 mentions) Murine thrombopoietin, by Thermo Fisher Scientific (3 mentions) Murine il 6, by Thermo Fisher Scientific (3 mentions) Human il 6, by Thermo Fisher Scientific (3 mentions) L glutamine, by Thermo Fisher Scientific (3 mentions) Erythropoietin, by Amgen (2 mentions) Human interleukin 6, by Thermo Fisher Scientific (2 mentions) Lineage cell depletion kit, by Miltenyi Biotec (2 mentions) X vivo 15 media, by Lonza (2 mentions) Erythropoietin, by Thermo Fisher Scientific (2 mentions) Il 11, by R&D Systems (2 mentions) Gm csf, by Thermo Fisher Scientific (2 mentions) Methocult 3434, by STEMCELL (2 mentions) Emu012611, by Merck Group (2 mentions) Ehuegfp, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

Murine stem cell factor cytokines Recombinant murine stem cell factor Recombinant murine stem cell factor (SCF;

29 protocols using murine stem cell factor

1

Inhibitors and Antibodies for Studying JAK-STAT Signaling

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The SFK PTK inhibitors Saracatinib (AZD0530) and SU6656 and the JAK PTK inhibitor CMP6 were purchased from Merck Millipore. Recombinant mouse IL-7, murine stem cell factor, IL-3, IL-6, and FLT3L were purchased from Peprotech. Antibodies against STAT5, p-(Y694) STAT5, p-(Y1022/1023) JAK-1, and JAK-1 were purchased from Cell Signaling. Tubulin was purchased from Sigma-Aldrich. Retronectin was purchased from Takara. Poly (I:C), Percoll, and Dnase I were from Sigma-Aldrich; FBS was from Thermo Fisher; Dulbecco-PBS (D-PBS) and HBSS were from Invitrogen; and collagenase D was from Roche. The mouse antibody against PTPN2 (6F3) and plasmid encoding murine PTPN2 (Ptpn2-pcDNA3.1) were provided by M. Tremblay (McGill University).
Wiede F., Dudakov J.A., Lu K.H., Dodd G.T., Butt T., Godfrey D.I., Strasser A., Boyd R.L, & Tiganis T. (2017). PTPN2 regulates T cell lineage commitment and αβ versus γδ specification. The Journal of Experimental Medicine, 214(9), 2733-2758.
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2

Fetal Liver Cell Colony Assay

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Lineage-depleted fetal liver cells were resuspended in MethoCult M3234 with cytokines supplemented at the following concentrations: erythropoietin (Amgen) 10 U/mL, murine stem cell factor (Peprotech) 50 ng/mL, murine IL-3 (Peprotech) 20 ng/mL, and murine IL-6 (Peprotech) 20 ng/mL. Cells were cultured at 37 degrees Celsius and colonies were counted at 3 and 7 days of culture. Colonies were visualized with a Nikon Eclipse TS100 microscope.
Li H., Natarajan A., Ezike J., Barrasa M.I., Le Y., Feder Z.A., Yang H., Ma C., Markoulaki S, & Lodish H.F. (2019). Rate of progression through a continuum of transit-amplifying progenitor cell states regulates blood cell production. Developmental cell, 49(1), 118-129.e7.
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3

Propagation of Murine MLL::AF9 AML Cells

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The murine MLL::AF9 AML model expressing Cas9 was previously generated.11 (link) Leukemia cells were serially propagated in sublethally irradiated C57BL/6 recipient mice, as previously described.11 (link)
MLL::AF9 leukemia cells were harvested from the bone marrow and enriched for LSCs by isolating the receptor tyrosine kinase+ (c-Kit+) cell population, and cultured in Stemspan (StemCell Technologies) containing 1% penicillin/streptomycin (Cytvia), supplemented with murine interleukin-3, murine stem cell factor, and human interleukin-6 (Peprotech).7 (link)
Rodriguez-Zabala M., Ramakrishnan R., Reinbach K., Ghosh S., Oburoglu L., Falqués-Costa A., Bellamkonda K., Ehinger M., Peña-Martínez P., Puente-Moncada N., Lilljebjörn H., Cammenga J., Pronk C.J., Lazarevic V., Fioretos T., Hagström-Andersson A.K., Woods N.B, & Järås M. (2023). Combined GLUT1 and OXPHOS inhibition eliminates acute myeloid leukemia cells by restraining their metabolic plasticity. Blood Advances, 7(18), 5382-5395.
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4

Establishing NOTCH1-induced T-ALL Model

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The NOTCH1-induced T-ALL model was established as previously decribed.17 (link) Briefly, bone marrow cells were isolated from 8-week-old donor mice. Lineage negative (Lin-) cells were enriched using a Lineage Cell Depletion Kit (Miltenyi Biotec), and pre-stimulated for 24 h in Dulbecco modified Eagle medium (Gibco) containing 20% fetal bovine serum (Gibco), 1% penicillin/streptomycin (Hyclone), 20 ng/mL murine FLT3-L, 20 ng/mL murine thrombopoietin and 100 ng/mL murine stem cell factor (PeproTech). Cells were then transduced with MigR1-ICN1 retroviruses and centrifuged in the presence of 6 Dg/mL polybrene. A second round of transduction was carried out on the next day. One million cells were then transplanted into sub-lethally (5.5 Gy) irradiated 8-week-old C57BL/6 female mice by tail vein injection.
Guo H., Xu J., Xing P., Li Q., Wang D., Tang C., Palhais B., Roels J., Liu J., Pan S., Huang J., Liu Z., Zhu P., Taghon T., Qing G., Van Vlierberghe P, & Liu H. (2023). RNA helicase DHX15 exemplifies a unique dependency in acute leukemia. Haematologica, 108(8), 2029-2043.
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5

Murine T-ALL Induction and Transplantation

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T-ALL was generated in mice as previously described (Herranz et al., 2015 ), with minor modifications. In brief, lineage negative hematopoietic cells were isolated from murine bone marrow using Miltenyi Lineage Cell Depletion Kit (130-090-858). Isolated cells were cultured for 16 hrs in X-VIVO 15 media (Lonza 04-418Q) with 100 ng/mL murine stem cell factor (Peprotech 250-03), 10 ng/mL murine IL3 (Peprotech 213-13), 10 ng/mL human IL6 (Peprotech 200-06) and 50 ng/mL FLT3-ligand (Peprotech 250-31L). Cells were then treated with freshly made MSCV-ICN1-IRES-GFP or MSCV-ICN1-IRES-NGFR or vector control virus along with 8 μg/mL polybrene on retronectin coated plates and centrifuged for 45 min at 1200 rpm. Cells were rested for 2 hr and plated with fresh media with cytokine supplements for 24 hrs. Spinfection protocol was repeated 24 hrs later. 48 hrs after final spinfection, 5 × 104 transduced cells (identified by GFP or NGFR) along with 2 × 105 bone marrow cells for hemogenic support were injected via tail vein into lethally irradiated (9.0 Gy split into two fractionated doses 4 hrs apart) syngeneic mice. Secondary recipient mice were irradiated with 4.5 Gy and received 2 × 105 T-ALL cells via tail vein.
Kishton R.J., Barnes C.E., Nichols A.G., Cohen S., Gerriets V.A., Siska P.J., Macintyre A.N., Goraksha-Hicks P., de Cubas A.A., Liu T., Warmoes M.O., Abel E.D., Yeoh A.E., Gershon T.R., Rathmell W.K., Richards K.L., Locasale J.W, & Rathmell J.C. (2016). AMPK is essential to balance glycolysis and mitochondrial metabolism to control T-ALL cell stress and survival. Cell metabolism, 23(4), 649-662.
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6

Directed Differentiation of iPSCs to Macrophages

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CD45.1 iPSCs were maintained in pluripotent state as described previously (Ackermann et al., 2014 (link)). Differentiation was performed after passaging the cells for 2 days in feeder-free conditions on a gelatin-treated plate in an IMDM medium (Thermo Fisher Scientific) containing 10% fetal calf serum (FCS; Millipore), 1 mM penicillin-streptomycin, 150 μM monothioglycerol, and 103 U/mL leukemia inhibitory factor (pre-culture medium). For embryoid body formation, single cells were seeded at a density of 10,000 cells/mL in differentiation medium I: IMDM medium with 15% pre-tested FCS (ES-Cult FBS, STEMCELL Technologies), 1 mM penicillin-streptomycin, 1 mM L-glutamine, 50 ng/mL ascorbic acid (Sigma-Aldrich), and 150 mM monothioglycerol (Sigma-Aldrich). After 5 days, the medium was supplemented with 10 ng/mL murine interleukin-3 and 30 ng/mL murine stem cell factor (PeproTech) (Mucci et al., 2016 (link), Pfaff et al., 2012 (link)). After 7 days, the embryoid bodies were dissociated into single cells using collagenase IV (250 U/mL, Life Technologies) and transferred into a suspension-culture plate with differentiation medium II: RPMI medium (Thermo Fisher Scientific) supplemented with 10% FCS, 1 mM L-glutamine, 1 mM penicillin-streptomycin, and 30% supernatant of L929 producer cells (Ladner et al., 1988 (link)) as a source of M-CSF.
Mucci A., Lopez-Rodriguez E., Hetzel M., Liu S., Suzuki T., Happle C., Ackermann M., Kempf H., Hillje R., Kunkiel J., Janosz E., Brennig S., Glage S., Bankstahl J.P., Dettmer S., Rodt T., Gohring G., Trapnell B., Hansen G., Trapnell C., Knudsen L., Lachmann N, & Moritz T. (2018). iPSC-Derived Macrophages Effectively Treat Pulmonary Alveolar Proteinosis in Csf2rb-Deficient Mice. Stem Cell Reports, 11(3), 696-710.
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7

Yolk Sac cKIT+ Cell Transfection

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E10.5 WT yolk sac (YS) cKIT+ cells (1×104) were plated in 100 μL maturation media (IMDM, 20% FBS, 1% interleukin-3 (IL-3) super-natant, 10 ng/ml murine stem cell factor (PeproTech), 10 ng/ml M-CSF (PeproTech), 10 ng/ml GM-CSF (PeproTech), 10 ng/ml IL-6, 10 ng/ml IL-11 (R&D Systems) and 2 U/ml erythropoietin (PeproTech) in a 96-well plate and transfected with 300 ng esiRNA against either Baf155 (Sigma, EMU012611) or Egfp (Sigma, EHUEGFP) with 2 μL lipofectamine 3000 (Thermofisher). Cells were cultured in a 37°C incubator with 5% CO2 for 36–48 hours and then subjected to either RNA extraction or re-plating in methylcellulose (MethoCult 3434, Stem Cell Technologies).
Wu J., Krchma K., Lee H.J., Prabhakar S., Wang X., Zhao H., Xing X., Seong R.H., Fremont D.H., Artyomov M.N., Wang T, & Choi K. (2020). Requisite Chromatin Remodeling for Myeloid and Erythroid Lineage Differentiation from Erythromyeloid Progenitors. Cell reports, 33(7), 108395.
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8

Hematopoietic Progenitor Assays

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Colony forming assays for megakaryocyte progenitors, erythroid and myeloid progenitors and high proliferative potential progenitors were performed as previously described (Palis et al., 2001 (link); Palis and Koniski, 2005 (link)). Embryonic tissues were explanted with two embryo equivalents in 100 ul of media as described (Shah et al., 2013 (link)). Sorted cells were cultured in maturation media (Arinobu et al., 2005 (link)) containing 20 ng/ml murine stem cell factor, IL-3, IL-6, and IL-7; 50 ng/ml IL-5 and IL-9; 10 ng/ml IL-11, granulocyte-macrophage colony-stimulating factor and thrombopoietin, (all Peprotech), and 2 U/ml erythropoietin (Amgen) at 105cells/ml or less. Short-term and long-term assays of B cell progenitors were performed as described (Yoshimoto et al., 2011 (link)).
McGrath K.E., Frame J.M., Fegan K.H., Bowen J.R., Conway S.J., Catherman S.C., Kingsley P.D., Koniski A.D, & Palis J. (2015). Distinct sources of hematopoietic progenitors emerge before HSCs and provide functional blood cells in the mammalian embryo. Cell reports, 11(12), 1892-1904.
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9

Coculture of HSCs with BMSCs

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2×105 lineage-negative cells (HSCs) were seeded over the BMSCs/M210B4 in IMDM supplemented with 20% Mesen-FBS (Stem Cell Technology), 25 ng/mL murine IL-6, 25 ng/mL murine stem cell factor and 10 ng/mL murine IL-3 (Peprotech, Rocky Hill, NJ), and the cocultures were maintained 7 days at 37°C under either normoxia (control-cocultures) or hypoxia (hypoxic-cocultures). Cocultures with CoCl2-BMSCs were incubated under normoxia (CoCl2-cocultures), unless stated otherwise.
After 7 days, the cocultures were harvested and viable cell counts were taken using the Trypan Blue dye exclusion method.
Moirangthem R.D., Singh S., Adsul A., Jalnapurkar S., Limaye L, & Kale V.P. (2015). Hypoxic Niche-Mediated Regeneration of Hematopoiesis in the Engraftment Window Is Dominantly Affected by Oxygen Tension in the Milieu. Stem Cells and Development, 24(20), 2423-2436.
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10

Mast Cell Differentiation from Murine Bone Marrow

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Mast cells were derived from bone marrow precursors by culturing stem cells flushed from tibia and femurs of 4-week-old female C57BL/6, Ahr−/− and Tph1−/− mice in RPMI 1640 supplemented with 20% fetal bovine serum (Euroclone), 2 mM glutamine (Sigma Aldrich), 100 U/ml penicillin streptomycin (Lonza), non essential amino acids (Sigma Aldrich), 1 mM sodium pyruvate (Sigma Aldrich), 20 mM HEPES (Sigma Aldrich), 5 ng/ml recombinant murine IL-3 (Peprotech) and 5 ng/ml murine stem cell factor (Milteniy) for 4 to 8 weeks. Cell cultures were grown at 37 °C in a humidified atmosphere with 5% CO2. Purity was measured by FACS by monitoring FcεRI and cKit receptor expression (> 90%).
Zelante T., Paolicelli G., Fallarino F., Gargaro M., Vascelli G., De Zuani M., Fric J., Laznickova P., Kohoutkova M.H., Macchiarulo A., Dolciami D., Pieraccini G., Gaetani L., Scalisi G., Trevisan C., Frossi B., Pucillo C., De Luca A., Nunzi E., Spaccapelo R., Pariano M., Borghi M., Boscaro F., Romoli R., Mancini A., Gentili L., Renga G., Costantini C., Puccetti M., Giovagnoli S., Ricci M., Antonini M., Calabresi P., Puccetti P., Di Filippo M, & Romani L. (2024). A microbially produced AhR ligand promotes a Tph1-driven tolerogenic program in multiple sclerosis. Scientific Reports, 14, 6651.
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