16 high sensitivity str kit

Cell line identity was validated by the MD Anderson Characterized Cell Line Core facility (supported by the NIH/NCI Cancer Center Support Grant, CA016672) with STR DNA fingerprinting, which used the Promega 16 High-Sensitivity STR Kit (Catalog # DC2100). The STR profiles were compared with online search databases (DSMZ/ATCC/JCRB/RIKEN) of 2455 known profiles and with the MD Anderson Characterized Cell Line Core database of 2556 known profiles. The KTC1 cell line STR profile was matched with a previously published profile [18 (link)].

All ovarian cancer cell lines (SKOV3ip1, HeyA8, OVCAR-8, OVCA-432) were maintained in RPMI 1640 media supplemented with 15% fetal bovine serum (FBS) and 0.1% gentamicin sulfate (GeminibBioproducts, Calabases, CA, USA; refs 47 (link), 48 (link)). The A498 and RCC4 cell lines were kindly provided by Dr Eric Jonasch (M.D. Anderson Cancer Center) and were maintained in DMEM-high glucose supplemented with 10% FBS and 0.1% gentamicin. The RF24 cells were maintained in MEM supplemented with 10% FBS, sodium pyruvate, MEM vitamins, L-glutamine, and MEM non-essential amino acids. 10T1/2 pericyte-like cells were maintained in DMEM-high glucose supplemented with 10% FBS and 0.1% gentamicin. All the cell lines were routinely tested to confirm the absence of Mycoplasma. Cell lines were validated by Short Tandem Repeat (STR) DNA fingerprinting using the Promega 16 High Sensitivity STR Kit. The STR profiles were compared with online search databases (DSMZ/ATCC/JCRB/RIKEN) of 2,455 known profiles; along with the MD Anderson Characterized Cell Line Core database of 2,556 known profiles. The STR profiles matched known DNA fingerprints or were unique. All the in vitro experiments were conducted with 60–80% confluent cultures. For cell growth assays, the cells were plated in six-well plates in triplicate and were counted each day for 6 days using a Beckman Coulter cell counter.

JJN3 cells were obtained from DSMZ (ACC 541). KMS11 cells were obtained from JCRB Cell Bank (JCRB1179). MM1R (CRL-2975), H929 (CRL-3580) and RPMI-8226 (CCL-155) cells were obtained from ATCC. Human bone marrow-derived mesenchymal stem cells (hMSC) were kindly provided by Dr. Michael Andreeff. Mycoplasma testing was routinely performed on all cell lines, and cell identity was validated by STR DNA fingerprinting using the Promega 16 High Sensitivity STR Kit. Primary BM samples from patients with MM relapsed disease after PI-based therapy and referred to the Department of Lymphoma and Myeloma at MD Anderson Cancer Center or the Department of Medicine and Surgery at the University of Parma were obtained after written informed consent with the approval of the institutions’ respective Institutional Review Boards (IRBs) and in accordance with the Declaration of Helsinki. The consent to publish the exact age and sex of the patients included in the study was obtained by the Department of Lymphoma and Myeloma at MD Anderson Cancer Center or the Department of Medicine and Surgery at the University of Parma. Patient characteristics are included in Supplementary Table 2. BM samples from healthy donors were obtained from AllCells.
The authors acknowledge the limitation of using MM cell lines to perform the experiments described in the manuscript.

Animal studies, including housing, care, euthanasia methods and experimental protocols, were approved by the University of Texas MD Anderson Cancer Center Animal Care and Use Committee, adhering to relevant guidelines. Two xenograft models were developed from patient‐derived tumours. The first (RMC2X) was derived from the same RMC primary tumour specimen as the cell line RMC2C using previously described methodology.³² A primary tumour sample was implanted orthotopically and subcutaneously and allowed to grow for three serial passages within SCID mice (Figure S1A). After the third passage, the xenograft tumour was compared to the primary tumour using STR DNA profiling to confirm the xenograft tumour maintained patient‐derived tumour tissue using the Promega 16 High Sensitivity STR Kit (catalogue #DC2100)²⁵ (Supporting Information S1). A second PDX model (RMC32X) was developed in a similar fashion. RMC32X was derived from the nephrectomy specimen of a black male patient with sickle cell trait who had received platinum therapy prior to nephrectomy; thus, the PDX served as a model of a platinum‐experienced tumour. Both primary tumours from which the two PDX models were derived have been previously molecularly characterised.⁸