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Cc100

Manufactured by STEMCELL
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Sourced in France, Canada

The CC100 is a multi-purpose centrifuge that can be used for a variety of laboratory applications. It features an adjustable speed range and a versatile rotor that can accommodate different types of sample tubes or plates. The CC100 provides consistent and reliable performance to meet the needs of various research and testing workflows.

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Lab products found in correlation

Methocult, by STEMCELL (3 mentions) Stemspan 2, by STEMCELL (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Erythropoietin, by Thermo Fisher Scientific (1 mentions) Erythropoietin, by Amgen (1 mentions) Stem cell factor (scf), by Thermo Fisher Scientific (1 mentions) Iscove s modified dmem, by Corning (1 mentions) Ab serum, by Atlanta Biologicals (1 mentions) Stemspan, by STEMCELL (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Insulin, by Merck Group (1 mentions) Dexamethasone (dex), by Merck Group (1 mentions) Polybrene, by Thermo Fisher Scientific (1 mentions) G csf, by Thermo Fisher Scientific (1 mentions) Gm csf, by Thermo Fisher Scientific (1 mentions) Hepes buffer, by Thermo Fisher Scientific (1 mentions) Facsaria 2, by BD (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Penicillin streptomycin, by STEMCELL (1 mentions) Easysep human cd34 positive selection kit, by STEMCELL (1 mentions)

Spelling variants of this product

StemSpan CC100 (24 citations) StemSpan CC100 cytokine cocktail (10 citations) CC100 cytokine cocktail (7 citations) Stem Span Cytokine Cocktail CC100 (2 citations)

16 protocols using cc100

1

MSC-HSPC Coculture Protocol

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After detachment from culture flasks, 5 × 103 MSCs were seeded in 40 μL medium per well of a Perfecta3D 96-well hanging drop plate (3D Biomatrix, Biotrend, Cologne, Germany). The developed spheroids were harvested and analyzed at different time points.
Coculture experiments were performed with a mixture of 5 × 103 MSCs and 5 × 102 CD34+ HSPCs per well in 40 μL medium consisting of GMP and serum-free expansion medium (SFEM; Stem Cell Technologies, Grenoble, France) at a 1 : 4 ratio supplemented with the recombinant human cytokines Flt-3 ligand, stem cell factor (100 ng/mL each), interleukin-3, and interleukin-6 (20 ng/mL each) (CC100; Stem Cell Technologies), from here on referred to as “GMP-SFEM-CC100 medium.” Cells were kept in a fully humidified atmosphere with 5% CO2 at 37°C. A partial medium exchange was performed every 2 to 3 days.
Schmal O., Seifert J., Schäffer T.E., Walter C.B., Aicher W.K, & Klein G. (2015). Hematopoietic Stem and Progenitor Cell Expansion in Contact with Mesenchymal Stromal Cells in a Hanging Drop Model Uncovers Disadvantages of 3D Culture. Stem Cells International, 2016, 4148093.
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2

Erythroid Differentiation of Expanded CD34+ Cells

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CD34+ cells were isolated from de-identified control patient peripheral blood leukoreduction filters. To limit variability from one individual to another, CD34+ isolated from 20 leukoreduction filters were pooled for each experiment. Blood components were separated using Ficoll-Opaque, and CD34+ cells were purified using anti-CD34 manual cell separation columns and conjugated microbeads according to manufacturer’s instructions (Miltenyi). Isolated peripheral blood CD34+ cells were expanded in H3000 media supplemented with CC100 (Stem Cell Technologies) for 4 days at a density of 105 cells/mL. CD34+ cells were differentiated toward erythrocytes using an adapted version of a previously described in vitro 3-phase culture system [20 (link)].Briefly, CD34+ cells were cultured in 3% (vol/vol) AB serum, 2% (vol/vol) human plasma, 10μg/mL insulin, 3U/mL heparin, 200μg/mL transferrin supplemented with either 10ng/mL stem cell factor (SCF), 1ng/mL interleukin-3 (IL-3) and 3IU/mL erythropoietin (EPO) (Phase I; D0–7) or 10ng/mL SCF and 3IU/mL EPO (Phase II; D7–11) or 1mg/Ml transferrin and 3IU/mL EPO (Phase III; D11–16). The treatments (0.1–10 μM pomalidomide, 10 μM HU, 0.1–10 μM compounds 3b, 3d, 3f and DMSO) were added to erythroid differentiation cultures at every culture medium change.
de Melo T.R., Dulmovits B., dos Santos Fernandes G.F., de Souza C.M., Lanaro C., He M., Al Abed Y., Chin C.M., Blanc L., Costa F.F, & dos Santos J.L. (2021). Synthesis and pharmacological evaluation of pomalidomide derivatives useful for Sickle Cell Disease treatment. Bioorganic chemistry, 114, 105077.
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3

Clonogenic Assay of Imatinib-Treated Cells

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Methylcellulose clonogenic assays were performed by plating cells (103) in 0.9% MethoCult (Stem Cell Technologies). Primary CD34+ cells were cultured with cytokines (CC100, Stem Cell Technologies) in the presence or absence of 1 µM imatinib and/or the indicated cytokines added directly to the medium. Cells were incubated in humid chambers at 37°C ± 0.1 µg/ml doxycycline in duplicate. Colonies were scored after 1–2 weeks in culture.
Gonzalez M.A., Olivas I.M., Bencomo‐Alvarez A.E., Rubio A.J., Barreto‐Vargas C., Lopez J.L., Dang S.K., Solecki J.P., McCall E., Astudillo G., Velazquez V.V., Schenkel K., Reffell K., Perkins M., Nguyen N., Apaflo J.N., Alvidrez E., Young J.E., Lara J.J., Yan D., Senina A., Ahmann J., Varley K.E., Mason C.C., Eide C.A., Druker B.J., Nurunnabi M., Padilla O., Bajpeyi S, & Eiring A.M. (2022). Loss of G0/G1 switch gene 2 (G0S2) promotes disease progression and drug resistance in chronic myeloid leukaemia (CML) by disrupting glycerophospholipid metabolism. Clinical and Translational Medicine, 12(12), e1146.
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4

Expansion and Evaluation of CD34+ Cells

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K562 (human myelogenous leukemia) cells
were purchased from ATCC. Cells were cultured in Iscove’s modified
Dulbecco’s medium containing 10% (v/v) fetal bovine serum and
5% penicillin and streptomycin, as previously described.53 (link) All cultures were grown at 37 °C in a humidified
environment containing 5% CO2.
CD34+ cells
were expanded in H3000 media supplemented with CC100 (Stem Cell Technologies)
for 4 days at a density of 105 cells/mL prior to use for
compound evaluation. Cells were treated for 24, 48, or 72 h with an
appropriate concentration of each analogue.
Holshouser S., Cafiero R., Robinson M., Kirkpatrick J., Casero RA J.r., Hyacinth H.I, & Woster P.M. (2020). Epigenetic Reexpression of Hemoglobin F Using Reversible LSD1 Inhibitors: Potential Therapies for Sickle Cell Disease. ACS Omega, 5(24), 14750-14758.
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5

Colony Formation Assay for Cytotoxicity

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Cells were treated, for 24 hours at a density of 1x105cells/mL, with the concentrations indicated for each drug or with vehicle (0.1% DMSO). Next, 1,250 (or 10,000 for primary CML) cells/well were seeded into a semisolid culture media consisting of 80% methylcellulose (MethoCult, STEMCELL Tech., Milan, Italy) and 20% RPMI-1640 (fully supplemented) or in methylcellulose medium supplemented with a cytokine cocktail (CC100; Stem Cell Technologies). After 6-10 days, colonies were counted using a phase-contrast microscope.
Riva B., Dominici M.D., Gnemmi I., Mariani S.A., Minassi A., Minieri V., Salomoni P., Canonico P.L., Genazzani A.A., Calabretta B, & Condorelli F. (2016). Celecoxib inhibits proliferation and survival of chronic myelogeous leukemia (CML) cells via AMPK-dependent regulation of β-catenin and mTORC1/2. Oncotarget, 7(49), 81555-81570.
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6

CD34+ Cell Expansion and Differentiation

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CD34+ expansion media: serum-free StemSpan supplemented with 10 mL/mL CC-100 (both Stemcell Technologies), 2 U/mL erythropoietin (Amgen), 10−6 M dexamethasone (Sigma) and 100 U/ml penicillin/streptomycin (GIBCO, ThermoFisher Scientific). Differentiation media: Iscove’s Modified DMEM (Cellgro) with 3% AB serum (Atlanta Biologicals), 2% human plasma (Stemcell Tech), 10 mg/mL insulin (Sigma), 3 U/mL heparin, 200 ug/mL transferrin (Athens Research & Technology), 10 ng/mL stem cell factor (Peprotech), 3 U/mL erythropoietin. Freeze media: 50% characterized fetal bovine serum (Hyclone), 10% dimethyl sulfoxide (Sigma), and 40% Iscove’s Modified DMEM. Thaw media: Iscove’s Modified DMEM supplemented with 5% characterized fetal bovine serum.
Dong A., Ghiaccio V., Motta I., Guo S., Peralta R., Freier S.M., Watt A., Damle S., Ikawa Y., Jarocha D., Chappell M., Stephanou C., Delbini P., Chen C., Christou S., Kleanthous M., Smith-Whitley K., Manwani D., Casu C., Abdulmalik O., Cappellini M.D., Rivella S, & Breda L. (2020). 2'-O-methoxyethyl splice-switching oligos correct splicing from IVS2-745 β-thalassemia patient cells restoring hemoglobin A production and chain rebalance. Haematologica, 106(5), 1433-1442.
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7

Transduction of Myeloid Progenitor Cells

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MNCs were cultured at up to 5x106 cells/mL in RPMI supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich, St. Louis, MO), 2 mM L-glutamine, 100 U/mL penicillin/streptomycin plus CC100 (StemCell Technologies, Vancouver, BC), GM-CSF (10 ng/mL), and G-CSF (10 ng/mL) (PeproTech, Rocky Hill, NJ). At 48 hours, cells were diluted to 106/mL and distributed into 6-well plates at 2 mL/well. shRNA lentiviral particles were added, targeting a multiplicity of infection (MOI) of 0.3, followed by the addition of polybrene (2 μg/mL) and HEPES buffer (Invitrogen, Carlsbad, CA; 10 mM). The cells were centrifuged at 1800 rpm for 90 minutes at 32°C. Following centrifugation, the plates were kept in a humidified incubator at 5% CO2 and 37°C. The medium was replaced with fresh medium at 12-18 hours after transduction. At 72 hours after transduction, the cells were pooled and RFP+ cells quantified by flow cytometry [Becton-Dickinson (BD) FACSAria II].
Yan D., Franzini A., Pomicter A.D., Halverson B.J., Antelope O., Mason C.C., Ahmann J.M., Senina A.V., Vellore N.A., Jones C.L., Zabriskie M.S., Than H., Xiao M.J., van Scoyk A., Patel A.B., Clair P.M., Heaton W.L., Owen S.C., Andersen J.L., Egbert C.M., Reisz J.A., D'Alessandro A., Cox J.E., Gantz K.C., Redwine H.M., Iyer S.M., Khorashad J.S., Rajabi N., Olsen C.A., O'Hare T, & Deininger M.W. (2021). SIRT5 Is a Druggable Metabolic Vulnerability in Acute Myeloid Leukemia. Blood Cancer Discovery, 2(3), 266-287.
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8

Expansion and Differentiation of CD34+ Cells

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CD34 + cells were isolated from cord blood using an EasySep human CD34-positive selection kit (STEMCELL Technologies, Canada). Cells were expanded by adding chemokine cocktails (CC100, STEMCELL Technologies, Canada) for three days, and the cells were then supplemented with recombinant human IL4 (50 ng/ml) and GM-CSF (50 ng/ml) cytokines and incubated at 37 °C and 5% CO2 for 5 additional days. Cell differentiation was characterized by staining for human CD11c.
Xu X., Petersen S., Rodriguez C, & Yi G. (2021). VISTA facilitates phagocytic clearance of HIV infected CEM-SS T cells. Heliyon, 7(7), e07496.
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9

Erythroid Differentiation of CD34+ HSPCs

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CD34+ HSPCs were thawed into a maintenance medium consisting of a StemSpan II base (StemCell Technologies), CC100 (StemCell Technologies), 50 ng/mL human TPO (Pepro Tech), and 1% penicillin/streptomycin (Life Technologies).61 (link),62 (link) Cells treated with RNP complexes for enhancer deletions were electroporated 48 hours after thawing and collected 72 hours post-nucleofection. Cells treated with lentivirus were transduced 24 hours after thawing, sorted 72 hours after thawing, and moved to erythroid media 96 hours after thawing.
After the maintenance phase, CD34+ HSPCs were differentiated using the three-phase culture system previously described.63 (link),64 (link) First, a base erythroid medium was created by supplementing IMDM with 2% human AB plasma, 3% human AB serum, 3 U/mL heparin, 10 μg/mL insulin, 200 μg/mL holo-transferrin, and 1% penicillin/streptomycin. From days 1-7 in erythroid media, this base medium was further supplemented with 3 U/mL EPO, 10 ng/mL human SCF, and 1 ng/mL IL-3. From days 7-12, this base medium was further supplemented with 3 U/mL EPO and 10 ng/mL human SCF. After day 12, the base medium was supplemented with 1 mg/mL total of holo-transferrin and 3 U/mL of EPO.
Cato L.D., Li R., Lu H.Y., Yu F., Wissman M., Mkumbe B.S., Ekwattanakit S., Deelen P., Mwita L., Sangeda R., Suksangpleng T., Riolueang S., Bronson P.G., Paul D.S., Kawabata E., Astle W.J., Aguet F., Ardlie K., de Lapuente Portilla A.L., Kang G., Zhang Y., Nouraie S.M., Gordeuk V.R., Gladwin M.T., Garrett M.E., Ashley-Koch A., Telen M.J., Custer B., Kelly S., Dinardo C.L., Sabino E.C., Loureiro P., Carneiro-Proietti A.B., Maximo C., Méndez A., Hammerer-Lercher A., Sheehan V.A., Weiss M.J., Franke L., Nilsson B., Butterworth A.S., Viprakasit V., Nkya S, & Sankaran V.G. (2023). Genetic regulation of fetal hemoglobin across global populations. medRxiv.
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10

Detailed Murine and Human Cell Cultures

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These mouse strains were purchased from The Jackson Laboratory (Sacramento, CA, USA): C57BL/6J, FVB/NJ, B6 (Cg)-Tyrc-2J Tg (UBC-mCherry) 1Phbs/J, FVB-Tg (CAG-luc,-GFP) L2G85Chco/J, and CByJ.B6-Tg (UBC-GFP) 30Scha/J. HEK293T and TC1 cells were cultured in DMEM (Invitrogen, Carlsbad, CA, USA) supplemented with 10% Fetal Calf Serum (HyClone, Chicago, IL, USA) and 1% penicillin and streptomycin (Invitrogen, Carlsbad, CA, USA). Expi293F cells were cultured in Expi293 Expression Media (Invitrogen, Carlsbad, CA, USA). Human CD34+ cells (AllCells, Alameda, CA, USA) were purchased and reported to be more than 96% pure. Murine bone marrow cells were maintained in StemSpan SFEM supplemented with CC100 (STEMCELL Technologies, Vancouver, BC, Canada), SFEM without supplement, or RPMI (Invitrogen, Carlsbad, CA, USA) with 1% Fetal Calf Serum (FCS). Animal protocols were approved by the Institutional Animal Care and Use Committee of The Scripps Research Institute (12-0029) or by the Institutional Ethics Committee and Institutional Animal Care Committee of the University of Ulsan College of Medicine (2016-02-168, 2017-12-281).
Han K.H., Arlian B.M., Lin C.W., Jin H.Y., Kang G.H., Lee S., Lee P.C, & Lerner R.A. (2020). Agonist Antibody Converts Stem Cells into Migrating Brown Adipocyte-Like Cells in Heart. Cells, 9(1), 256.
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