molm 14 cell lines  (DSMZ)

Journal: EMBO Reports

Article Title: AML patient blasts exhibit polarization defects upon interaction with bone marrow stromal cells

doi: 10.1038/s44319-025-00466-w

Figure Lengend Snippet: ( A ) Schematic drawing representing microwells on a cover slide. The first zoom illustrates the non-adhesive edges (polyacrylamide) and the adhesive bottom (fibronectin coating), allowing the attachment and confining of stromal cells. The second magnification illustrates the position of the centrosome in the hematopoietic cell relative to cell contact with the stromal cell and the measurement of the polarization index (d/D). ( B ) Immunostainings of the centrosome (red), the nucleus (blue) and the actin cytoskeleton (green) in Cord blood (CB) HSPC and AML cell lines interacting with osteoblasts (hFOB) within microwells. Images were acquired in 3D. The top row shows a top projected view and scale bars correspond to 10 µm. Bottom row shows a side projected view and scale bars correspond to 10 µm. CB HSPCs form a magnupodium in contact with stromal cells. MOLM-14 and NOMO-1 leukemic blasts are both round and do not exhibit a magnupodium upon interaction with the osteoblast. ( C ) Graphs show the quantification of the polarization index of AML cell lines, MOLM-14 and NOMO-1, in contact with osteoblasts (hFOB) in comparison to CB HSPCs within microwells. Experiments were repeated two or three times and data were pooled ( n CB HSPCs = 59, n MOLM-14 = 95; n NOMO-1 = 173). Each spot corresponds to the measurement of the polarity of a single cell. The various shapes of the spots correspond to distinct experiments. Black bars show the median value and the standard deviation of the distribution. Differences between populations were evaluated using non-parametric test Kruskal–Wallis ANOVA with a P value < 0.0001 (****).

Article Snippet: MOLM-14 AML cell line , ATCC , #CRL-3006.

Techniques: Adhesive, Comparison, Standard Deviation

Journal: EMBO Reports

Article Title: AML patient blasts exhibit polarization defects upon interaction with bone marrow stromal cells

doi: 10.1038/s44319-025-00466-w

Figure Lengend Snippet: ( A ) The microfluidic circuit of the BMOC comprises 6 parallel channels. Channels (1) and (2) (not shown) are committed to medium supplementation. Channels (3), (4) and (5) are dedicated to endosteal (hFOBs), endothelial (HUVECs) and fibroblastic cells (NHLFs), respectively. All stromal cells are cultured in an hydrogel composed of collagen and fibrin. Endosteal compartment harbors central pillars to resist hydrogel contraction due to osteoblasts traction forces. The central channel (6) is devoted to loading hematopoietic cells such as hematopoietic stem and progenitor cells (HSPCs), AML cell lines (MOLM-14 or NOMO-1), or AML patient blasts. The photography on the left shows the BMoC perfused with colorants to illustrate the position of the distinct compartments. Microscopy images on the right show centrosome, actin filaments and nucleus stainings within the endosteal and endothelial compartments. Scale bars correspond to 50 µm. ( B ) Immunostainings of the centrosome (red, pointed at with white arrowheads), the nucleus (blue) and the actin cytoskeleton (green) of cord blood (CB) HSPCs and AML cell lines, MOLM-14 and NOMO-1, interacting with osteoblasts (hFOBs) in the endosteal compartment. Scale bars correspond to 10 µm. ( C ) Quantification of the polarization index of CB HSPCs (red dots), MOLM-14 and NOMO-1 (blue dots) in contact with osteoblasts. Experiments were repeated three to five times and data were pooled ( n CB HSPCs = 240, n MOLM-14 = 81; n NOMO-1 = 153). Each spot corresponds to the measurement of the polarity of a single cell. The various shapes of the spots correspond to distinct experiments. Black bars show the median value and the standard deviation of the distribution. Differences between populations were evaluated using Kruskal–Wallis ANOVA test with P values of 0.0002 and <0.0001 (***,****). ( D ) Same as ( B ) with CB HSPCs, MOLM-14 and NOMO-1 interacting with endothelial cells (HUVECs) in the BMoC endothelial compartment. Scale bars correspond to 10 µm. ( E ) Same as ( C ) with CB HSPCs, MOLM-14 and NOMO-1 interacting with endothelial cells (HUVECs) in the BMoC endothelial compartment. Experiments were repeated three times and data were pooled ( n CB HSPCs = 106, n MOLM-14 = 167; n NOMO-1 = 215). Differences between populations were evaluated using Kruskal–Wallis ANOVA test with P values < 0.0001 and 0.0008 (****,***).

Article Snippet: MOLM-14 AML cell line , ATCC , #CRL-3006.

Techniques: Cell Culture, Microscopy, Standard Deviation

Journal: EMBO Reports

Article Title: AML patient blasts exhibit polarization defects upon interaction with bone marrow stromal cells

doi: 10.1038/s44319-025-00466-w

Figure Lengend Snippet: ( A ) AML cell line (MOLM-14) and AML patient blasts (blue in the schematics) were loaded in the BMOC and thus interacted with MSC from healthy donors (orange in the schematics). Images in transmitted light were extracted from time-lapse movies of AML cell lines and patient blast within healthy compartments. Scale bars correspond to 50 µm. ( B ) Quantification of the mean speed and traveled distance during 20 min of the migration of leukemic cells (MOLM-14) or AML patient blast in a healthy stromal compartment (HD MSC). Tracks were analyzed at various positions within the stromal compartment ( n MOLM-14/ HD MSC = 41, n AML blast/ HD MSC = 77). In the violin plots, black bars represent the median and dashed lines the 95% confidence interval. Differences between populations were evaluated using a Mann–Whitney test with P values < 0.0001 (****).

Article Snippet: MOLM-14 AML cell line , ATCC , #CRL-3006.

Techniques: Migration, MANN-WHITNEY

Journal: Cell death & disease

Article Title: High iASPP (PPP1R13L) expression is an independent predictor of adverse clinical outcome in acute myeloid leukemia (AML).

doi: 10.1038/s41419-024-07190-8

Figure Lengend Snippet: Fig. 4 Functional analysis of iASPP in a MOLM14 interference cell line model. A, B Transduction efficiency assessed by (A) mRNA assessed by RT-qPCR and (B) protein levels assessed by flow cytometry. C iASPP-interference results in reduced metabolic activity as assessed by XTT, D corresponding to an attenuated proliferation rate with a prolonged cellular doubling time (16 h for iASPP KD vs. 13 h for iASPP EV). E, F, G Induction of apoptosis in response to daunorubicin, cytarabine or FLT3/multikinase-inhibitor sunitinib. H, I, J Box plot analysis to determine IC50 concentrations compared to empty vector (EV) control strains. Technical triplicates minimum for all assays. *p ≤0.05, **p ≤0.01, ≤***p ≤0.001, ****p ≤0.0001 (t-test).

Article Snippet: Mycoplasma-certified leukemia cell lines MOLM14 and HL60 were obtained from the German Collection of Microorganisms and Cell Cultures (Leipniz Institute DSMZ, Germany).

Techniques: Functional Assay, Transduction, Quantitative RT-PCR, Cytometry, Activity Assay, Plasmid Preparation, Control

Journal: Cell death & disease

Article Title: High iASPP (PPP1R13L) expression is an independent predictor of adverse clinical outcome in acute myeloid leukemia (AML).

doi: 10.1038/s41419-024-07190-8

Figure Lengend Snippet: Fig. 5 Apoptosis signaling of MOLM14 iASPP-interferenced vs. EV cell strains in response to daunorubicin. A Heat map of a proteome profiler human apoptosis array covering 35 proteins involved in apoptosis control. B Upregulation of proteins involved in induction of apoptosis and cell cycle control in the iASPP knock down (KD) vs EV cell strains.

Article Snippet: Mycoplasma-certified leukemia cell lines MOLM14 and HL60 were obtained from the German Collection of Microorganisms and Cell Cultures (Leipniz Institute DSMZ, Germany).

Techniques: Control, Knockdown

Journal: Cell death & disease

Article Title: High iASPP (PPP1R13L) expression is an independent predictor of adverse clinical outcome in acute myeloid leukemia (AML).

doi: 10.1038/s41419-024-07190-8

Figure Lengend Snippet: Fig. 6 Xenotransplant mouse model according to iASSP expression. A Transduction efficiency assessed by mRNA. B In vivo visualization of luciferase activity in MOLM14_iASPP KD_Luc+ vs. MOLM14_EV_Luc+ xenotransplanted mice on day 26 post transplantationem. 3 representative mice shown. C Luciferase activity over time. D Survival rates over time (n = 4 EV / 4 KD). E Whole femur of a representative MOLM14 EV control mouse, stained with DAPI, tdTomato, Endomucin, aSMA and CollagenIV, 10x magnification. F, G Zoomed-in images of BM regions that were densely infiltrated by MOLM14 cells. (H) MOLM14-iASPP KD Luc+ mouse femur with a single MOLM14 infiltration zone under the growth plate of the distal epiphysis and (I, J) zoomed-in image of the proximal epiphysis. **p ≤0.01 (Log-Rank Mantel Cox test), ***p ≤0.001 (unpaired t-test).

Article Snippet: Mycoplasma-certified leukemia cell lines MOLM14 and HL60 were obtained from the German Collection of Microorganisms and Cell Cultures (Leipniz Institute DSMZ, Germany).

Techniques: Expressing, Transduction, In Vivo, Luciferase, Activity Assay, Control, Staining

Journal: Cell Death & Disease

Article Title: High iASPP (PPP1R13L) expression is an independent predictor of adverse clinical outcome in acute myeloid leukemia (AML)

doi: 10.1038/s41419-024-07190-8

Figure Lengend Snippet: A Transduction efficiency assessed by mRNA. B In vivo visualization of luciferase activity in MOLM14_ iASPP KD_Luc+ vs. MOLM14_EV_Luc+ xenotransplanted mice on day 26 post transplantationem. 3 representative mice shown. C Luciferase activity over time. D Survival rates over time ( n = 4 EV / 4 KD). E Whole femur of a representative MOLM14 EV control mouse, stained with DAPI, tdTomato, Endomucin, aSMA and CollagenIV, 10x magnification. F , G Zoomed-in images of BM regions that were densely infiltrated by MOLM14 cells. ( H ) MOLM14- iASPP KD Luc+ mouse femur with a single MOLM14 infiltration zone under the growth plate of the distal epiphysis and ( I , J ) zoomed-in image of the proximal epiphysis. ** p ≤ 0.01 (Log-Rank Mantel Cox test), *** p ≤ 0.001 (unpaired t-test).

Article Snippet: Mycoplasma-certified leukemia cell lines MOLM14 and HL60 were obtained from the German Collection of Microorganisms and Cell Cultures (Leipniz Institute DSMZ, Germany).

Techniques: Transduction, In Vivo, Luciferase, Activity Assay, Control, Staining