Rpmi 1640

VCR-R CEM and CCRF-CEM cells were obtained from Prof. Maria Kavallaris (University of New South Wales, Sydney, Australia) [21 (link)] and were cultured in RPMI 1640 (PAN Biotech, Aidenbach, Germany) containing 10% FCS (PAN Biotech). VCR-R CEM cells are a multidrug-resistant subline of CCRF-CEM cells [22 (link)], generated by long-term exposure to increasing concentrations of vincristine [23 (link)]. Jurkat cells were bought from ATCC and cultured in RPMI 1640 containing 10% FCS and pyruvate. Cell lines were typically passaged twice to thrice a week. HeLa cells were purchased from DSMZ (Braunschweig, Germany) and cultured in DMEM (PAN Biotech) containing 10% FCS. The model of ALL patients’ leukemia cells growing in mice has been described previously [24 (link)]. In the present study, patient-derived xenograft (PDX) cells were engrafted and freshly isolated from the bone marrow or spleen of NSG mice (The Jackson Laboratory, Bar Harbour, ME, USA) and subsequently cultured in RPMI 1640 supplemented with 20% FCS and P/S. Peripheral blood mononuclear cells (PBMC) were obtained from ATCC and cultured in RPMI 1640 supplemented with 20% FCS and P/S.
Cell line STR profiling was performed. None of the cell lines used are listed in the database of commonly misidentified cell lines maintained by ICLAC. All cells are proven to be mycoplasma-free quarterly.

Human umbilical vein endothelial cells (HUVEC, ATCC CRL-1730) were grown in DMEM (Lonza) with FBS (10%, PAA), ultraglutamine (1%, Lonza) and gentamicin sulfate (Lonza) at 37 °C and CO₂ (5%). Murine alveolar MH-S macrophages (ATCC CRL-2019) were grown in RPMI 1640 (Lonza) supplemented with heat-inactivated FBS (10% FBS), sodium-pyruvate (1 mM, Lonza), ultraglutamine (1%) and gentamicin sulfate (50 mg mL⁻¹) at 37 °C and CO₂ (5%). THP-1 cells (DSMZ, ATCC 16) were maintained in RPMI 1640 supplemented with FBS (10%), ultraglutamin (2 mM) and gentamicine sulfate at 37 °C and CO₂ (5%). U-937 cells (ATCC CRL-1593.2) were cultered in RPMI 1640 supplemented with FBS (10%) and gentamicin sulfate at 37 °C and CO₂ (5%).
Cells were authenticated and tested for mycoplasma contamination by ATCC, passaged every second day until passage 30. Normal human serum (NHS) was prepared from FHR1/3 sufficient as well as FHR1/3 deficient (ΔFHR1/3 NHS) blood derived from healthy volunteers, as determined by PCR^{52 (link)} and Western blot analyses. After coagulation blood was centrifuged (10 min, 2000×g, 4 °C), and NHS kept frozen in aliquots at −80 °C. C. albicans cph1Δ/efg1Δ^{53 (link), 54 (link)}, which cannot form hyphae, was grown overnight in YPD medium (D glucose (2%), peptone (1%), yeast extract in H₂O (5%) at room temperature.

All cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA). T47D clones (CTL and SPEN) were stably transfected with control and SPEN-expressing vectors as previously described and were cultured in Gibco DMEM (Life Technologies, Carlsbad, CA, USA) supplemented with 10% FBS (Wisent Bio Products, Saint-Bruno, PQ, Canada) and G418 (500 μg/ml) under conditions specified by the manufacturer. BT20 and MDA-MB-436 cells were cultured in Eagle’s minimum essential medium and RPMI 1640 (ATCC), respectively, supplemented with 10% FBS. MCF10A cells were maintained in RPMI 1640 (ATCC) containing 10% FBS, 10 μg/ml insulin, 0.5 μg/ml hydrocortisone, and 20 ng/ml human epidermal growth factor. MCF10A-CT cells were maintained in Gibco DMEM/Ham’s F-12 Nutrient Mixture (Life Technologies) containing 5% horse serum, 10 μg/ml insulin, 0.5 μg/ml hydrocortisone, 20 ng/ml human epidermal growth factor, and 100 ng/ml cholera toxin (CT).