Rpmi8226

Four MM cell lines (MM.1S, RPMI8226, KMS11, and U266) and monocytic osteoclast progenitor cell line (RAW264.7) were purchased from Procell (Wuhan, China). MM cell lines and RAW264.7 cells were incubated in RPMI-1640 or DMEM medium (Meilunbio, China), which contained 10% FBS (ExCell, China), penicillin and streptomycin. 5% CO₂ at 37 °C was used to culture all cells. From September 2019 to January 2021, paraffin sections of bone marrow samples of 24 MM patients and 7 iron deficiency anemia (IDA) patients were obtained from the First Affiliated Hospital of Fujian Medical University. And the percentage of CD138 + cells in the bone marrow of MM patients was obtained from the pathology report. Primary plasma cells were isolated from bone marrow specimens of patients with MM and healthy donors using anti-CD138 MicroBeads (Miltenyi, Germany) and immediately frozen in − 80 °C until the subsequent extraction of RNA. The Ethics Committee of the First Affiliated Hospital of Fujian Medical University approved this study. According to the Declaration of Helsinki, informed consent was obtained.

Human MM cell lines of RPMI8226 and KMS11 were purchased from Procell ( Catalogue number: CL-0564 and CL-0794). MM cell lines were maintained in RPMI 1640 medium containing 10% FBS (Supplier name: ExCell, China;Catalogue number: FND500), penicillin and streptomycin at 37 ℃ in a humidified atmosphere of 5% CO₂ atmosphere. Cells were monitored for mycoplasma contamination using a mycoplasma detection kit (Supplier name:Beyotime; Catalogue number: C0301S). RPMI8226 and KMS11 cells stably transfected with NCX1-overexpress lentivirus (established by Shanghai HANBIO Company, China) or NCX1-shRNA lentivirus (established by Shanghai Genechem Company, China) were named RPMI8226-oeNCX1 / KMS11-oeNCX1 or RPMI8226-shNCX1 / KMS11-shNCX1, respectively. The control cell lines named RPMI8226-oeCON / KMS11-oeCON or RPMI8226-shCON / KMS11-shCON with the same empty vectors were developed in the same way.

HEK293T cells were cultured in DMEM (Hyclone®). RPMI-8226 from Procell (Wuhan, China) and LP1, MM1.S and MM1.R maintained in the lab were cultured in IMDM (Hyclone®), all were added with 10% FBS (ExCell Bio, Inc., Shanghai, China) and 1% penicillin/streptomycin (Sigma-Aldrich®). Primary bone marrow cells were donated by healthy donors and MM patients from the First Affiliated Hospital of Soochow University with written consent for research purposes and approved by the Review Board and Ethic Committee of Soochow University.