This study was approved by the institutional review board of The University of Texas MD Anderson Cancer Center (Houston, TX, USA). ARP-1 and ARK cells were kindly provided by Arkansas Cancer Research Center (AR, Usa). Others were purchased from the American Type Culture Collection (ATCC). Primary myeloma cells were isolated from bone marrow aspirates of myeloma patients using anti-CD138 antibody-coated magnetic beads (Miltenyi Biotec, Inc. San Diego, CA, USA). Recombinant human resistin was purchased from PeproTech (Rocky Hill, NJ, USA). Except where specified, all chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA), all antibodies for flow cytometry analysis were purchased from BD Biosciences (San Jose, CA, USA), and all antibodies for western blot analysis were purchased from Cell Signaling Technology (Danvers, MA, USA). The short interfering RNA (siRNA) against human ABCC5 and ABCG2 genes as well as the non-target control siRNA were purchased from Santa Cruz Technologies (Dallas,TX, USA).
Anti cd138 antibody coated magnetic beads
Manufactured by Miltenyi Biotec
Sourced in Germany
Anti-CD138 antibody-coated magnetic beads are a laboratory product designed for the isolation and enrichment of CD138-positive cells from biological samples. The beads are coated with monoclonal antibodies specific to the CD138 surface antigen, which is commonly expressed on plasma cells and some types of cancer cells. These magnetic beads can be used in various cell separation and purification applications.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi8226, by American Type Culture Collection (3 mentions)
Hek293t, by American Type Culture Collection (2 mentions)
Recombinant human resistin, by Thermo Fisher Scientific (1 mentions)
Rpmi 8226 ccl 155, by American Type Culture Collection (1 mentions)
Nci h929, by American Type Culture Collection (1 mentions)
Arh 77, by American Type Culture Collection (1 mentions)
Mm 1s, by American Type Culture Collection (1 mentions)
8 protocols using anti cd138 antibody coated magnetic beads
1
Isolation and culture of primary myeloma cells
2
Myeloma Cell Lines and Patient Samples
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Myeloma cell lines ARP-1, H929 and U266 were provided by Dr. Zhiqiang Liu's lab of Tianjin Medical University. The MCF7 (#HTB-22), HEK293T (#ACS-4500), RPMI8226 (CCL-155), and MM.1S (CRL-2974) cells were purchased from the American Type Culture Collection. Primary myeloma cells were isolated from bone marrow aspirates of myeloma patients by using anti-CD138 antibody-coated magnetic beads (Miltenyi Biotec). Myeloma cells were maintained in RPMI1640 medium with 10% fetal bovine serum (FBS). MCF7 and HEK293T cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% FBS. Patient samples were obtained from the Qingdao Municipal Hospital of Qingdao University and The First Affiliated Hospital of Xiamen University. Bone lesions in the study participants were characterized by radiologists at Qingdao Municipal Hospital. This study was approved by the Ethics Committee of Xiamen University, and all protocols conformed to the Ethical Guidelines of the World Medical Association Declaration of Helsinki.
3
Myeloma Cell Lines and Patient Samples
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The myeloma cell line ARP-1 was provided by the University of Arkansas for Medical Sciences (Little Rock, Arkansas, USA). HEK293T and RPMI8226 cells were purchased from the American Type Culture Collection (Manassas, VA, USA). Primary myeloma cells were isolated from bone marrow aspirates from patients with myeloma using anti-CD138 antibody-coated magnetic beads (Miltenyi Biotec, Bergisch Gladbach, Germany). Myeloma cells were maintained in RPMI 1640 medium with 10% fetal bovine serum (FBS), and HEK293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS. Patient samples were obtained from UT MD Anderson Cancer Center and Biorepository of Houston Methodist Research Institute. Bone lesions in the study participants were characterized by radiologists from Bone Survey at UT MD Anderson Cancer Center. The radiologists were blinded to the severity of clinical disease. This study was approved by the Institutional Review Boards of UT MD Anderson Cancer Center and Houston Methodist Research Institute.
4
Procurement and Culture of Myeloma Cells
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Human myeloma cell lines were purchased from American Type Culture Collection (ATCC), except that ARP-1 and ARK cells were kindly provided by Arkansas Cancer Research Center, AR. The p53 knockout myeloma cells and drug resistant myeloma cells against bortezomib, dexamethasome, or carfilzomib were established as previously described before [11] (link). Primary myeloma cells were isolated from bone marrow aspirates of myeloma patients using anti-CD138 antibody-coated magnetic beads (Miltenyi Biotec, Inc.). This study was approved by the institutional review board of The University of Texas MD Anderson Cancer Center (Houston, TX). All myeloma cells were cultured in RPMI 1640 medium (Mediatech Cellgro) supplemented with 10% fetal bovine serum (FBS) and antibiotics.
5
Isolation and Culture of Primary MM Cells
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ARP-1 and ARK cells were kindly provided by Arkansas Cancer Research Center, AR. Others were purchased from American Type Culture Collection (ATCC). Primary MM cells were isolated from BM aspirates of MM patients using anti-CD138 antibody-coated magnetic beads (Miltenyi Biotec, Inc.). Cells were maintained in RPMI1640 medium with 10% fetal bovine serum (FBS). This study was approved by the institutional review board of The University of Texas MD Anderson Cancer Center (Houston, Texas).
6
Isolation of Primary Myeloma Cells
7
Isolation and Culture of Multiple Myeloma Cells
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RPMI8226, ARH-77, U266, NCI-H929 and 3T3-L1 cells were purchased from American Type Culture Collection (ATCC). Primary MSCs were isolated from BM aspirates of healthy control volunteers using anti-CD138 antibody coated magnetic beads (Miltenyi Biotec, Inc.). Cells were cultured in conditioned medium with 10% fetal bovine serum (FBS) according to the ATCC cell culture suggestion. Multiple myeloma cells were grown in RPMI-medium containing 2 mmol/L glutamine, 20% FCS, 100 U/mL penicillin and 0.1 mg/mL streptomycin.
8
Isolation and Culture of Multiple Myeloma Cells
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The MM cell line ARP-1 was provided by the University of Arkansas for Medical Sciences. Murine MM Vk*MYC cell line (Vk12598) was provided by the Mayo Clinic [20 (link)]. HEK293T, MM.1S, U266, and RPMI8226 cells were purchased from the American Type Culture Collection. Primary MM cells were isolated from bone marrow aspirates from patients with MM using anti-CD138 antibody-coated magnetic beads (Miltenyi Biotec, Germany). MM cells were maintained in RPMI 1640 medium with 10% fetal bovine serum (FBS), and HEK293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS. All patient samples were obtained from the Biorepository of Houston Methodist Research Institute (HMRI) or the Myeloma Tissue Bank of UT MD Anderson Cancer Center (MDACC). This study was approved by the Institutional Review Board of HMRI and MDACC. In some experiments, MM cells were cultured with 20 μg exosomes/105 cells or treated with chemotherapeutic drugs, such as bortezomib (5 nM), melphalan (25 μM), or carfilzomib (25 nM).
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