In vitro model | Type of model | Culture condition | Source (identifier) |
AML | |||
Kasumi-1 | Cell lines | Standard RPMI | Princess Máxima Center (CVCL_0589) |
SKNO1 | RPMI+Glutamax supplemented with 20% FCS, 1% P/S and 10 ng/mL GM-CSF (Miltenyi Biotec). | Princess Máxima Center (CVCL_2196) | |
PDX01 | PDX | PDX material derived from primary t(8;21) pediatric acute myeloid leukemia15 was cocultured with human bone-marrow derived MSCs as feeder layer. MSCs were cultured in low glucose DMEM (Life Technologies) supplemented with 20% FCS, 8 ng/mL FGF-2 (PeproTech) and 1% P/S and were seeded in 24 well plate at a density of 7500 cell/cm2 1 day prior to PDX culture. PDX cells were then thawed and resuspended at 1×106 cell/mL in an in-house optimized SFEMII medium (STEMCELL Technologies) supplemented with cytokines including 150 ng/mL SCF, 100 ng/mL TPO, 10 ng/mL FLT3-L, 10 ng/mL IL-3 (all PeproTech), 1350 nM UM729 (STEMCELL Technologies), 750nM SR1 (Biogems), and 10 ng/mL GM-CSF (Princess Máxima Center pharmacy). Cells were kept on MSCs for 3 days before cells were harvested and functional T-cell assays were performed. | Princess Máxima Center |
B-ALL | |||
NALM6 | Cell Lines | Standard RPMI | Princess Máxima Center (CVCL_0092) |
RCH-ACV | Princess Máxima Center (CVCL_1851) | ||
SUPB-15 | Princess Máxima Center (CVCL_0103) | ||
697 | Princess Máxima Center (CVCL_0079) | ||
T-ALL | |||
MOLT16 | Cell lines | Standard RPMI | Princess Máxima Center (CVCL_1424) |
CCRF-CEM | Princess Máxima Center (CVCL_0207) | ||
HCC | |||
pHCC | Organoid | The culture medium was based on Saltsman et al.16 with modifications as reported in17 . The medium consisted of the following: Advanced DMEM/F12 supplemented with 1% P/S, 1% GlutaMAX, 10 mM HEPES, 1 x B27 supplement, 1x N2 supplement (all Life Technologies), 0.2% Normocin (Invivogen), 1.25 mM N-acetyl-L-cysteine, 10% (vol/vol) RSPO1 conditioned medium, 10 mM nicotinamide, 10 nM recombinant human (Leu15)-gastrin I (All Sigma), 50 ng/mL recombinant human EGF, 25 ng/mL recombinant human HGF (Both Peprotech), 10 µM forskolin, 5 µM A8301 (Both Tocris Bioscience), 10 µM Y27632 ((Abmole), and 0.5 nM WNT surrogate.18 Cell culture and passing were performed as described by Kluiver et al.19. Basement membrane extract (BME) (R&D systems) 3D-cultures were dissociated using dispase and TripleE (both Life Technologies). Cells were washed and resuspended in the culture medium above + 2% FCS. Plates were coated with collagen I (Corning) and cells were cultured in 2D-setting before assays were initiated. | Princess Máxima Center |
NBL | |||
GIMEN | Cell lines | DMEM+Glutamax supplemented with 10% FCS, 1% P/S, 2% non-essential amino acids (Life Technologies). | Amsterdam Medical Center (CVCL_1232) |
IMR32 | Princess Máxima Center (CVCL_0346) | ||
CHP212 | RPMI+Glutamax supplemented with 10% FCS, 1% P/S, 2% non-essential amino acids. | Dana Farber Institute (CVCL_1125) | |
SMSKAN | Dana Farber Institute (CVCL_7131) | ||
SKNDZ | Dana Farber Institute (CVCL_1701) | ||
NLF | Dana Farber Institute (CVCL_E217) | ||
691b | Organoids | NBL organoids were established, characterized and cultured as described previously20. Organoids were maintained in DMEM low glucose, Glutamax-supplemented medium, supplemented with 25% HAM’s F-12 Nutrient Mix, 2% B-27 supplement minus vitamin A, 1% N-2 Supplement, 1% P/S (all Life Technologies), 20 ng/mL Animal-Free Recombinant Human-EGF, 40 ng/mL FGF-basic, 200 ng/mL IGF-I, 10 ng/mL PDGF-AA, and 10 ng/mL PDGF-BB (all Peprotech). | Princess Máxima Center |
039 | |||
MRT | |||
78T2 | Organoids | MRT organoids were established, characterized, and cultured as described previously21 22. In short, organoids were grown in droplets of growth factor-reduced BME type 2 (R&D Systems), advanced DMEM/F12 containing 1X GlutaMAX, 10 mM HEPES, and 1% P/S, supplemented with 10% R-spondin–conditioned medium, 1.5% B27 supplement, 50 ng/mL EGF, 50 ng/mL FGF-2 (PeproTech), 1.25 mM N-acetylcysteine, 10 mM ROCK inhibitor Y-27632, and 5 mM A83-01. Cells were cultured in 2D-setting in functional T-cell assays. | Princess Máxima Center |
JD081T | |||
JD041T | |||
60T2 | |||
HBL | |||
HB13F | Organoids | HBL organoids were established, characterized, and cultured as described previously17, see HCC for detailed culture conditions. | Princess Máxima Center |
HB13E | |||
DMG | |||
JHH-DMG-01 | Neurospheres | Neurospheres were established, characterized, and cultured as previously described23 24. Cells were maintained in DMEM/F12 and Neurobasal-A medium (1:1), supplemented with, 0.2% Primocin, 10% FCS, 1M HEPES, 1% non-essential amino acids, 1× Glutamax, 1% sodium pyruvate, 2% B-27 supplement (without vitamin A) (All Life Technologies), 20 ng/mL bFGF, 20 ng/mL EGF, 10 ng/mL PDGF-AA, 10 ng/mL PDGF-BB (All Preprotech), and 5 IE/mL Heparin (Pharmacy PMC). Biweekly, a half medium change was performed and once a week Accutase (400–600 units/mL) was used to disrupt spheres to circumvent overcrowding and necrotic core formation. | Princess Máxima Center |
HSJD-DMG-07 | |||
Adult Cancer Controls | |||
K562 | Cell lines | Standard RPMI | UMC Utrecht (CVCL_0004) |
Jurkat | UMC Utrecht (CVCL_0065) |
Standard RPMI=RPMI + Glutamax (Life Technologies), supplemented with 10% FCS (Sigma)+1% Penicillin/Streptomycin (P/S) (Life Technologies).
ALL, acute lymphoblastic leukemia; AML, acute myeloid leukemia; DMG, diffuse midline glioma; HBL, hepatoblastoma; HCC, hepatocellular carcinoma; MRT, malignant rhabdoid tumor; MSCs, mesenchymal stem cells; NBL, neuroblastoma; PDX, patient-derived xenograft.