cll cell line mec1  (DSMZ)

Journal: Cell Reports Methods

Article Title: Dynamic stimulation promotes functional tissue-like organization of a 3D human lymphoid microenvironment model in vitro

doi: 10.1016/j.crmeth.2025.101105

Figure Lengend Snippet: The disease model (A) Recirculation of MEC1 CLL cells in the “niche” conformation with the microenvironment. Immunofluorescence images showing CD45 + (green) MEC1 cells in the LN and BM microenvironments. Z Stacks were acquired using a 20× (a, d), a 30× (c), a 60× (e, f), and a 60× with 1.5 zoom (b) objective lens. Scale bars, 50 μm. (B) Flow cytometry analysis of CLL-specific markers on circulating cells in the LN model. (C) Flow cytometry analysis of CLL-specific markers on circulating cells in the BM model. For all the markers, data are represented by fold change relative to the 2D condition. All data are represented by mean ± SEM. For each model (LN and BM) two independent experiments (EXP.1 in black, EXP.2 in pink), with four replicates each were performed.

Article Snippet: The CLL cell line MEC1 was obtained from DSMZ (Cat. No. ACC 497; DSMZ, Braunschweig, Germany).

Techniques: Immunofluorescence, Flow Cytometry

Journal: Cell death & disease

Article Title: IL-4-dependent Jagged1 expression/processing is associated with survival of chronic lymphocytic leukemia cells but not with Notch activation.

doi: 10.1038/s41419-018-1185-6

Figure Lengend Snippet: Fig. 1 Jag1 is constitutively processed in CLL cells. a–c Western blot analysis of Jag1 was performed in 25 µg whole-cell lysates (WCL) from primary CLL cells (n = 21), MEC1 cell line, and PBL from healthy donors (n = 3), using three different C-terminal antibodies (Abs), as indicated on top of the blots. Data in panel a are obtained using the Jag1 C-20 Ab. Data in panels b and c are from representative CLL samples where the analysis of Jag1 was repeated using the Jag1 TS1.15H (b) and Jag1 E-12 (c) Abs. WCL isolated from IM9 cell line was used as a positive control for Jag1 expression. Protein loading was assessed reprobing the blots with an anti-GAPDH antibody. In each blot, separated panels are shown because different X-ray film exposures were necessary to detect Jag1-FL and Jag1 fragments. Positions of the molecular weight markers (kDa) are indicated on the right of the blots. In the blots with PBL, an empty lane has been placed between PBL2 and CLL11 to avoid cross-contamination. Vertical line inserted in the middle blot of panel b indicates a repositioned gel lane. Full images of all blots are shown in Supplementary Figure S1

Article Snippet: Normal PBL were isolated from healthy donors as previously reported.23 The human CLL MEC1 cell line was obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany), and cultured in Iscove’s MDM containing 10% fetal bovine serum.

Techniques: Western Blot, Isolation, Positive Control, Expressing, Molecular Weight

Journal: Cell death & disease

Article Title: IL-4-dependent Jagged1 expression/processing is associated with survival of chronic lymphocytic leukemia cells but not with Notch activation.

doi: 10.1038/s41419-018-1185-6

Figure Lengend Snippet: Fig. 2 Jag1 processing generates an extracellular fragment released into CLL culture medium and an intracellular fragment that translocates into the nucleus. a Western blot analysis of sJag1-EC was performed, using the N-terminal Jag1 AF1277 antibody, in soluble proteins (40 µg) of conditioned medium (CM) collected from CLL cells (n = 10; CLL4-13, selected to include patients with different clinical and biological characteristics), and from MEC1 cell line cultured for the indicated times in serum-free conditions. Serum-free medium (indicated as medium) incubated in the same conditions of CLL and MEC1 cultures, but without leukemic cells, was used as negative control. WCL from each respective CLL sample and from MEC1 cell line were used as a control for the molecular weight of Jag1-FL. Ponceau red staining is shown to assess protein loading. Representative CLL cases are shown. Full images of the blots are shown in Supplementary Figure S2A. b Western blot analysis of Jag1 was performed, using the C-terminal Jag1 C-20 antibody, in 25 μg nuclear (NE) and cytoplasmic (Cyt) extracts from CLL (n = 10; CLL4-13) and MEC1 cells. Adequate fractionation and protein loading were assessed using anti-lamin B1 and anti-β-tubulin antibodies. WCL from each respective CLL sample and from MEC1 cells were used as a control for the molecular weight of Jag1-TM and Jag1-IC. An empty lane was placed between WCL, NE and Cyt lanes to avoid potential cross-contamination. Representative CLL cases are shown. Full images of the blots are shown in Supplementary Figure S2B. c Confocal microscopy images of subcellular localization of Jag1-IC in a representative CLL sample and in MEC1 cells. CLL cells (n = 5; CLL7-11) and MEC1 cells were stained with the C-terminal Jag1 HPA021555 antibody (green) and with DAPI for nuclei (blue), as described in “Materials and methods”, and then analyzed by confocal microscopy. Upper images show a representative field, lower images show one single slice from the z-stacks of a representative CLL or MEC1 cell in the field. Scale bars are indicated in each image

Article Snippet: Normal PBL were isolated from healthy donors as previously reported.23 The human CLL MEC1 cell line was obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany), and cultured in Iscove’s MDM containing 10% fetal bovine serum.

Techniques: Western Blot, Cell Culture, Incubation, Negative Control, Control, Molecular Weight, Staining, Fractionation, Confocal Microscopy

Journal: Cell reports

Article Title: Targeting Macrophages Sensitizes Chronic Lymphocytic Leukemia to Apoptosis and Inhibits Disease Progression.

doi: 10.1016/j.celrep.2016.01.042

Figure Lengend Snippet: Figure 1. BM Microenvironment of Xeno- transplanted Mice: Monocyte and Macro- phage Populations (A–F) Rag2/gc/ mice, uninjected (UN, black circles) or injected i.v. with MEC1 (white circles) cells (day 0), were killed at early (n = 3) and late (n = 3) stage of leukemia and analyzed by flow cy- tometry (A). The mean value of the relative contri- bution of hCD19+ cells in BM is shown in (B). The mean value of the absolute number of CD11b+

Article Snippet: The MEC1 CLL cell line was obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) and cultured in RPMI 1640medium (Invitrogen) with 10% fetal bovine serum and gentamicin (15 mg/ml; SigmaAldrich).

Techniques: Injection

Journal: Cell reports

Article Title: Targeting Macrophages Sensitizes Chronic Lymphocytic Leukemia to Apoptosis and Inhibits Disease Progression.

doi: 10.1016/j.celrep.2016.01.042

Figure Lengend Snippet: Figure 2. Effect of Clodrolip-Mediated Macrophage Depletion in CLL Transplantation Systems (A–D) Rag2/gc/ mice, injected i.v. with MEC1 cells (day 0) and macro- phage-depleted by i.v. injection of PBS liposomes (200 ml, black circles, C) or clodrolip (200 ml, white circles, clo) every 3 days starting at day 1 were sacrificed at day 28 (A). Histology of spleen is shown (one representative mouse): H&E staining and immunohistochemistry stain for murine F4/80; scale bars, 100 mm (B). Spleens from control and clodrolip-treated mice were collected and analyzed macroscopically; for control mice, half the spleen is shown (C). The mean value of the relative contribution of hCD19+ cells in SP, BM, PB, and PE is shown in graph (D). Data are from two independent

Article Snippet: The MEC1 CLL cell line was obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) and cultured in RPMI 1640medium (Invitrogen) with 10% fetal bovine serum and gentamicin (15 mg/ml; SigmaAldrich).

Techniques: Transplantation Assay, Injection, Liposomes, Staining, Immunohistochemistry, Control

Journal: Cell reports

Article Title: Targeting Macrophages Sensitizes Chronic Lymphocytic Leukemia to Apoptosis and Inhibits Disease Progression.

doi: 10.1016/j.celrep.2016.01.042

Figure Lengend Snippet: Figure 3. Monocyte/Macrophage Killing Approach in CLL Xenograft Systems at the Preclinical Level (A–G) Rag2/gc/ mice transplanted s.c. with MEC1 cells were treated s.c. at the LN site (days +39, +45, +52, +56, +60, +63, and +66) with 50 ml of PBS liposomes (n = 5, black circles) or clodrolip (n = 5, white circles) (A). A representative mouse (day 48) with its tumor (dashed line) and LN (continuous line) growth curves are shown for PBS liposomes (B and C) and clodrolip (D and E). Tumor and LN volumes (mean ± SD) are shown (F). Statistical analysis was performed using the Student’s t test (*p < 0.05, **p < 0.01, and ***p < 0.001; measurements were stopped when 60% of mice were still surviving). These two groups of mice were monitored for survival (G). Kaplan-Meier survival curve is shown, statistical analyses was performed using the log-rank test (***p < 0.001). Data are from one representative experiment of two. (H and I) i.v. MEC1 challenged (day 0) Rag2/gc/ mice were treated i.v. (days +10, +14, +17, and +20) with 60 ml PBS liposomes (n = 3, black circles) or clodrolip (n = 3, white circles) and killed at day 32 (H). The mean value of the absolute number of hCD19+ cells in the BM is shown in graph (I). Statistically significant differences were calculated using the Student’s t test (**p < 0.01).

Article Snippet: The MEC1 CLL cell line was obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) and cultured in RPMI 1640medium (Invitrogen) with 10% fetal bovine serum and gentamicin (15 mg/ml; SigmaAldrich).

Techniques: Liposomes

Journal: Cell reports

Article Title: Targeting Macrophages Sensitizes Chronic Lymphocytic Leukemia to Apoptosis and Inhibits Disease Progression.

doi: 10.1016/j.celrep.2016.01.042

Figure Lengend Snippet: Figure 4. Anti-leukemic Effect of Anti-CSF1R mAb in the CLL Xeno-transplantation System (A–E) i.v. MEC1 challenged (day 0) Rag2/gc/ mice were treated i.v. (days +11 and +25) with 30 mg/kg anti-CSF1R mAb (n = 3, white circles) or left untreated (n = 4, black circles) and killed at day 27 (A). The mean value of the absolute number of CD11b+ F4/80+ cells gated on CD45+ in SP (B) and BM (C) is shown. The mean value of the absolute number of human CD19+ cells in the BM is shown (D). The mean value of the absolute number of human CD19+ cells and of AnnVPI+ cells gated on hCD19+ cells in SP is shown (E). Data are from one representative experiment of three. Statistical analysis: *p < 0.05 and **p < 0.01, Student’s t test. (F–I) i.v. MEC1 challenged (day 0) Rag2/gc/ mice were treated i.v. (days +11 and +25) with 30 mg/kg anti-CSF1R mAb (n = 7, white circles) or left untreated (n = 7, black circles) and killed at days 27 and 29, 48 hr and 96 hr after the last mAb injection (F). The mean value of the absolute number of human

Article Snippet: The MEC1 CLL cell line was obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) and cultured in RPMI 1640medium (Invitrogen) with 10% fetal bovine serum and gentamicin (15 mg/ml; SigmaAldrich).

Techniques: Transplantation Assay, Injection

Journal: Cell reports

Article Title: Targeting Macrophages Sensitizes Chronic Lymphocytic Leukemia to Apoptosis and Inhibits Disease Progression.

doi: 10.1016/j.celrep.2016.01.042

Figure Lengend Snippet: Figure 5. Anti-leukemic Effect and Survival Impact of Anti-CSF1R mAb in the CLL Xeno-transplantation System (A–C) i.v. MEC1 challenged (day 0) Rag2/gc/

Article Snippet: The MEC1 CLL cell line was obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) and cultured in RPMI 1640medium (Invitrogen) with 10% fetal bovine serum and gentamicin (15 mg/ml; SigmaAldrich).

Techniques: Transplantation Assay

Journal: Cell reports

Article Title: Targeting Macrophages Sensitizes Chronic Lymphocytic Leukemia to Apoptosis and Inhibits Disease Progression.

doi: 10.1016/j.celrep.2016.01.042

Figure Lengend Snippet: Figure 6. TNF-Dependent Macrophage- Mediated Mechanism of In Vivo Leukemic Cell Death (A–G) i.v. MEC1 challenged (day 0) Rag2/gc/

Article Snippet: The MEC1 CLL cell line was obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) and cultured in RPMI 1640medium (Invitrogen) with 10% fetal bovine serum and gentamicin (15 mg/ml; SigmaAldrich).

Techniques: In Vivo