Seven cHL cell lines (L-428, HDLM-2, KM-H2, L-1236, U-HO1, SUP-HD1, L-540) and 10 NHL cell lines (RAJI, DAUDI, RAMOS, NAMALWA, CA-46, VAL, OCI-LY1, OCI-LY3, OCI-LY7, SU-DHL-6) were obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ) or were kindly provided by Andreas Bräuninger (University Hospital Giessen, Germany) (cells: L-428, KM-H2, L-1236, L-540) [12 (link),13 (link)]. Detailed information on cell culture conditions are presented in Table S1 . Cell lines were authenticated by STR DNA profiling.
Sudhl6
Manufactured by Leibniz Institute DSMZ
Sourced in Germany
The SUDHL6 is a specialized cell line derived from a human diffuse large B-cell lymphoma. It is commonly used in research applications to study the biology and potential therapeutic targets of this type of lymphoma. The core function of the SUDHL6 cell line is to provide a model system for in vitro experiments and analyses related to this particular disease.
Automatically generated - may contain errors
Lab products found in correlation
Su dhl 4, by Leibniz Institute DSMZ (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Rpmi 1640, by Thermo Fisher Scientific (4 mentions)
Karpas 422, by Leibniz Institute DSMZ (3 mentions)
Oci ly7, by Leibniz Institute DSMZ (2 mentions)
Oci ly3, by Leibniz Institute DSMZ (2 mentions)
U2932, by Leibniz Institute DSMZ (2 mentions)
Su dhl 10, by Leibniz Institute DSMZ (2 mentions)
Dohh 2, by Leibniz Institute DSMZ (2 mentions)
Sudhl8, by Leibniz Institute DSMZ (2 mentions)
Mec 1, by Leibniz Institute DSMZ (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Namalwa, by Leibniz Institute DSMZ (1 mentions)
L 428, by Leibniz Institute DSMZ (1 mentions)
Hdlm 2, by Leibniz Institute DSMZ (1 mentions)
Km h2, by Leibniz Institute DSMZ (1 mentions)
L1236, by Leibniz Institute DSMZ (1 mentions)
L 540, by Leibniz Institute DSMZ (1 mentions)
Oci ly1, by Leibniz Institute DSMZ (1 mentions)
12 protocols using sudhl6
1
Cell Line Authentication and Culture Protocols
2
DLBCL Cell Line Cultivation and SHP-1 Agonist Assay
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The DLBCL cell lines, DB cells were obtained from the American Type Culture Collection (Manassas, VA, USA). U2932, SU-DHL-6, and OCI-Ly7 cells were purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen (Braunschweig, Germany). U2932, SU-DHL-6 and DB cells were maintained in RPMI 1640 Medium supplemented with 20%, 20%, and 10% fetal bovine serum (FBS) respectively. OCI-Ly7 cells were cultured in Iscove's Modified Dulbecco’s Medium supplemented with 20% FBS. All cell lines were incubated at 37 °C in a 5% CO2 incubator. SHP-1 agonists, SC-43 and SC-60, were synthesized, and its quality was evaluated as described in a previous study (Liu et al. 2017a (link), 2013 (link)). For cell-based studies, SHP-1 agonists were dissolved in dimethyl sulfoxide (DMSO) and then added to the cells in medium containing 5% FBS. The Myc-DDK-tagged Lyn expression construct and pCMV6-Entry plasmids were purchased from OriGene (Rockville, MD, USA). DLBCL cells were transiently transfected with the PolyJet transfection reagent (SignaGen Laboratories, Frederick, MD, USA) following to manufacturer’s instructions. To knockdown endogenous SHP-1, cells were transfected with siRNAs against PTPN6 (L-009778-00) or control (D-001810-10) (final concentration of 25 μM) for 72 h using DharmaFECT 1 Transfection Reagent (T-2001-01) according to manufacturer's instructions (Dharmacon, Chicago, IL, USA).
3
DLBCL Cell Line Viability Assay
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The human GCB-like DLBCL cell lines, SUDHL4, and SUDHL6, were obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ, Braunschweig, Germany) and the two ABC-like DLBCL cell lines, OCI-LY3, and OCI-LY10, were a gift of Pr Feuillard (UMR CNRS 7276, Limoges University) with the kind agreement of Louis M Staudt (National Cancer Institute, USA). The cell lines were grown in RPMI 1640 medium (Lonza, Basel, Switzerland) supplemented with 10% heat-inactivated fetal bovine serum (Thermo Scientific HyClone, Logan, UT, USA), 100 U ml−1 penicillin, 100 μg ml−1 streptomycin (Gibco Invitrogen Corporation, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere containing 5% CO2. For the viability tests, cells were incubated with: various concentrations of rituximab (1–20 μg ml−1, MabThera, stock 10 mg ml−1, a generous gift from CHRU Dupuytren of Limoges, Pharmacie centrale), K252a (350 nM ; Alomone Labs, Jerusalem, Israel), pro-BDNF (1–10 ng ml−1, Alomone Labs), neutralising anti-BDNF (15 μg ml−1, Promega, Madison, WI, USA), exogenous BDNF (100 ng ml−1, Promega), anti-p75NTR neutralising antibody (1–10–100 ng ml−1, clone ME20.4, Merck KGaA, Darmstadt, Germany) alone or in combination.
4
Cell Line Generation and Cultivation Protocol
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DOHH2, SUDHL4 (FL), SUDHL6, SUDHL10, OCILY3, Karpas 422 and SUDHL5 (DLBCL) were obtained from DSMZ (Braunschweig, Germany). DOHH2, SUDHL4, OCILY3, Karpas 422 cells were routinely grown at 37°C at 5% CO2 in RPMI 1640 supplemented with 10% fetal calf serum (FCS), ultra-glutamine, penicillin and streptomycin (100U/ml). SUDHL5, SUDHL6 and SUDHL10 cells were cultured with 20% FCS. Five LCLs were generated from peripheral blood mononuclear cells by infection with B95.8 virus. One LCL was used to compare in the 2-D experiments, the other four were used in the validation and functional studies. LCLs were routinely grown in RPMI 1640 with 10% FCS. For the production of LCLs from peripheral blood permission was granted by the Institutional Review board (medical ethical committee UMCG) and written informed consent was obtained.
5
Characterization of DLBCL Cell Lines
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Human DLBCL cell lines Karpas-422, SU-DHL-4, WSU-DLCL2, SU-DHL-10, SU-DHL-6, DB, DOHH-2, HT, and SU-DHL-8 were purchased from DSMZ. The TOLEDO cell line was purchased from ATCC. OCI-LY1 and OCI-LY3 were provided by the Louis M. Staudt lab (Center for Cancer Research, National Cancer Institute, NIH, Bethesda, Maryland, USA); HBL-1 and OCI-LY10 were provided by the Michael Gold lab (Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada). Karpas-422, SU-DHL-4, SU-DHL-10, SU-DHL-8, SU-DHL-6, and DB were cultured in RPMI 1640 (Thermo Fisher Scientific) supplemented with 20% FBS (Thermo Fisher Scientific). WSU-DLCL2, DOHH-2, OCI-LY1, HT, HBL-1, and TOLEDO were cultured in RPMI 1640 supplemented with 10% FBS. OCI-LY1 and OCI-LY10 were cultured in IMDM supplemented with 10% and 20% FBS, respectively. All cell lines were confirmed to be negative for mycoplasma using Venor GeM Mycoplasma Detection Kit, PCR-based (MilliporeSigma, MP0025). All cell lines were authenticated by STR profiling (The Centre for Applied Genomics, The Hospital for Sick Children, Toronto, Canada, Supplemental Table 10 ). Mutations in EZH2 in the cell lines were evaluated using the COSMIC (https://cancer.sanger.ac.uk/cosmic ) databases and in-house whole-genome or whole-exome sequencing data.
6
Chronic B-cell Leukemia Cell Lines
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Chronic B-cell leukemia-derived cell lines MEC-1 (RRID: CVCL_1870), EHEB (RRID: CVCL_1194), and lymphoma lines SUDHL6 (RRID: CVCL_2206), BJAB (RRID: CVCL_5711), and Bal17 (RRID: CVCL_9474) were purchased from DSMZ (Braunschweig, Germany). MEC-1 (Slc7a) Eco cells were generated by transduction with pLenti6/UbC/mSlc7a1, a gift from Shinya Yamanaka (Addgene plasmid # 17224; RRID: Addgene_17224) and MEC-1 Luciferase positive cells were generated by the introduction of pCL6-Luc-GFP into MEC-1 Eco cells by retroviral transduction.
7
Lymphoma Cell Line Maintenance Protocol
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Burkitt’s lymphoma (Glor, BL31, Sav, Ezema, Dante) and diffuse large B cell lymphoma (Farage, SUDHL4, SUDHL5, SUDHL6) were purchased from DSMZ (Braunschweig, Germany). Cell lines were maintained in exponential growth in RPMI-1640 media (Gibco-Invitrogen Ltd., Paisley, UK) supplemented with 10% fetal bovine serum (FBS, Gibco-Invitrogen), penicillin (100 U/mL) and streptomycin (100 μg/mL) (Gibco-Invitrogen) at 37 °C with 5% CO2. The cultures were routinely passaged every 2 days to maintain exponential phase. Cells were authenticated regularly with the NorthGene service for STR profiling. The mycoplasma test was performed with DAPI stain (Sigma Aldrich, St. Louis, MO, USA).
8
PBMC Isolation, Cell Purification, and Activation Protocols
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Primary peripheral blood mononuclear cells (PBMC) were isolated by Ficoll separation (Ficoll-Paque, GE Healthcare). Normal CD19 + B cells and CD4 + T cells were prepared from human PBMC by B Cell Isolation Kit II and CD4 + T Cell Isolation Kit, respectively (Miltenyi Biotec). The immunopurified cells were confirmed by flow cytometry. DLBCL cell lines SUDHL4, SUDHL6, and SUDHL8 were purchased from DSMZ. Adult T cell leukemia (ATL) patient-derived TL-Om1 cell line was a gift from Dr. K. Sugamura, Tohoku University, Japan. B95.8 EBV-transformed lymphoblastoid cell lines (LCLs) were previously established by infection of lymphocytes from four independent healthy donors with culture supernatants of the virus producer B95.8 line56 (link). All lymphoid cell lines and primary lymphocytes were cultured in RPMI1640 (GIBCO) supplemented with 10% of FBS (GIBCO) and antibiotics (GIBCO). 293T, 293FT, and MDA-MB-231 cells were maintained in DMEM (Nissui, Japan) with 10% of FBS and antibiotics.
B cell activation was performed by anti-IgM (Jackson Immunoresearch Laboratories), BAFF (R&D Systems), anti-CD40 (R&D Systems), and/or IL-4 (R&D Systems). T cell activation was performed by anti-CD3/CD28 antibodies (Miltenyi Biotec).
B cell activation was performed by anti-IgM (Jackson Immunoresearch Laboratories), BAFF (R&D Systems), anti-CD40 (R&D Systems), and/or IL-4 (R&D Systems). T cell activation was performed by anti-CD3/CD28 antibodies (Miltenyi Biotec).
9
Umbilical Cord Blood Collection Protocol
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CB was obtained postpartum from umbilical cords of full-term healthy babies after normal vaginal delivery by cannulation of the umbilical vein using the method we described previously in the National Heart, Lung and Blood Institute for Cord Blood Transplantation study [25] . Briefly, after aseptic preparation of the cord, the umbilical vein was punctured using a sterile 18 gauge needle attached to sterile tubing on a 200 mL sterile collection bag according to the manufacturer's directions (Pall Medical, Port Washington, NY, USA). The bag contained 25 mL of citrate phosphate dextrose as an anticoagulant. CB was collected by gravity flow with constant rocking. All samples were collected after approval by the institutional review board and were performed in compliance with the Declaration of Helsinki.
Cell lines and tumor targets NK sensitive, K562 (human chronic myelogenous leukemia) (ATCC, Manassas, VA, USA), NK-resistant Daudi (human Blymphoblastic cell line, ATCC), Ramos (BL, ATCC), and SUDHL-6 (diffuse large B cell lymphoma [DLBCL], DSMZ, Braunschweig, Germany) cell lines were cultured and maintained in RPMI-1640 (Invitrogen, Carlsbad, CA, USA) with 10% heatinactivated fetal bovine serum (FBS, Invitrogen) at 37 C and 5% CO 2 .
Cell lines and tumor targets NK sensitive, K562 (human chronic myelogenous leukemia) (ATCC, Manassas, VA, USA), NK-resistant Daudi (human Blymphoblastic cell line, ATCC), Ramos (BL, ATCC), and SUDHL-6 (diffuse large B cell lymphoma [DLBCL], DSMZ, Braunschweig, Germany) cell lines were cultured and maintained in RPMI-1640 (Invitrogen, Carlsbad, CA, USA) with 10% heatinactivated fetal bovine serum (FBS, Invitrogen) at 37 C and 5% CO 2 .
10
Establishment and Characterization of DLBCL Cell Lines
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