Journal: Biology Open

Article Title: Development of a highly pulmonary metastatic orthotopic renal cell carcinoma murine model

doi: 10.1242/bio.058566

Figure Lengend Snippet: Bioluminescent imaging (BLI) of orthotopically implanted Renca cells and spontaneous lung metastases. (A) Representative bioluminescent images of BALB/c mice following intrarenal injection of 1×10 5 Renca/luc or Renca(HM)/luc cells. (B) Higher BLI signal intensity (higher tumor burden) on day 5 following Renca(HM)/luc cell implantation (relative to day 9 following Renca/luc cell implantation) ( n =2 for each day 5, 9, 14, 19, and 23 post implantation, total n =10 for Renca/luc and Renca(HM)/luc, respectively).

Article Snippet: To induce stable expression of red fluorescent protein (RFP) and firefly luciferase (luc), Renca cells were transfected using a commercially available lentiviral vector containing luc 3 and RFP genes under control of an inducible suCMV promoter (Amsbio, Cambridge, MA, USA).

Techniques: Imaging, Injection

Journal: Biology Open

Article Title: Development of a highly pulmonary metastatic orthotopic renal cell carcinoma murine model

doi: 10.1242/bio.058566

Figure Lengend Snippet: Schematic representation of the in vivo selection process. Cell lines were intrarenally injected (orthotopic implantation) in mouse (A), followed by isolation of tumor cells from pulmonary metastases for expansion culture and reinjection in another mouse (B). (1) Initial Renca/luc cell implantation. (2) Isolation of metastatic tumor cells from pulmonary lesions. (3) In vivo selection of the highly pulmonary metastatic cell subpopulation, termed Renca(HM)/luc. (4) Final reimplantation of Renca(HM)/luc cells.

Article Snippet: To induce stable expression of red fluorescent protein (RFP) and firefly luciferase (luc), Renca cells were transfected using a commercially available lentiviral vector containing luc 3 and RFP genes under control of an inducible suCMV promoter (Amsbio, Cambridge, MA, USA).

Techniques: In Vivo, Selection, Injection, Isolation

Journal: Biology Open

Article Title: Development of a highly pulmonary metastatic orthotopic renal cell carcinoma murine model

doi: 10.1242/bio.058566

Figure Lengend Snippet: Progression rate of primary renal tumors and lung metastatic lesions. (A) Kaplan–Meier survival curves, indicating significantly shorter survival intervals for Renca(HM)/luc cell-implanted mice than for Renca/luc cell-implanted mice ( n =10 for Renca/luc and Renca(HM)/luc). The dotted line represents the Kaplan–Meier survival curve following final Renca(HM)/luc cell-reimplantation(2nd study, n =10). (B) Excised tumor-bearing kidney weights of Renca/luc cell-implanted mice relative to those of Renca(HM)/luc cell-implanted mice ( n =10 for Renca/luc tumor-bearing, and Renca(HM)/luc tumor bearing kidneys). (C) Higher weight of the tumor-bearing kidney (more rapid tumor growth) following Renca(HM)/luc cell implantation (relative to Renca/luc cell implantation) ( n =2 for each day 5, 9, 14, 19, and 23 post-implantation, total n =10 for Renca/luc and Renca(HM)/luc, respectively).

Article Snippet: To induce stable expression of red fluorescent protein (RFP) and firefly luciferase (luc), Renca cells were transfected using a commercially available lentiviral vector containing luc 3 and RFP genes under control of an inducible suCMV promoter (Amsbio, Cambridge, MA, USA).

Techniques:

Journal: Biology Open

Article Title: Development of a highly pulmonary metastatic orthotopic renal cell carcinoma murine model

doi: 10.1242/bio.058566

Figure Lengend Snippet: Spontaneous lung metastasis following intrarenal implantation of highly pulmonary metastatic Renca(HM)/luc cells. (A) Bioluminescent imaging demonstrating early (day 5) lung metastatic lesions in Renca(HM)/luc cell-implanted mice versus no signal in Renca/luc cell-implanted mice. (B) Number of pulmonary metastatic lesions over time, demonstrating more lesions in Renca(HM)/luc cell-implanted mice than in Renca/luc cell-implanted mice. (C) Pulmonary metastatic lesions visualized via India ink-insufflation in Renca/luc cell-implanted mice and Renca(HM)/luc cell-implanted mice on day 23 post-implantation. Scale bars: 5 mm. (D) Number of pulmonary metastases over time, demonstrating significantly more lesions in Renca(HM)/luc cell-implanted mice than in Renca/luc cell-implanted mice [ n =2 for each day 5, 9, 14, 19, and 23 post-implantation, total n =10 for Renca/luc and Renca(HM)/luc, respectively].

Article Snippet: To induce stable expression of red fluorescent protein (RFP) and firefly luciferase (luc), Renca cells were transfected using a commercially available lentiviral vector containing luc 3 and RFP genes under control of an inducible suCMV promoter (Amsbio, Cambridge, MA, USA).

Techniques: Imaging

Journal: Oncology Letters

Article Title: Anti-4-1BB antibody-based combination therapy augments antitumor immunity by enhancing CD11c + CD8 + T cells in renal cell carcinoma

doi: 10.3892/ol.2021.13161

Figure Lengend Snippet: Combined therapy with ISTF and anti-4-1BB mAb has antitumor effects in mice inoculated with Renca cells. Mice were inoculated with Renca tumor cells on day 0. Tumor-bearing mice were divided into four groups and treated with the following reagents: i) PBS and control mAb (rat IgG); ii) ISTF monotherapy; iii) 4-1BB mAb monotherapy; and iv) ISTF and 4-1BB mAb combined. Mice were subcutaneously injected with Renca tumor cells (1×10 6 /mouse). ISTF (100 µg/mouse) was injected i.p. on days 3, 7 and 12, and anti-4-1BB mAb (100 µg/mouse) was injected i.p. on days 7 and 12, while a control group received PBS and/or rat IgG, respectively. Tumor diameter was measured every 2–3 days, and tumor volume (in mm 3 ) was calculated using a caliper. Tumor size was expressed as tumor volume based on the following formula: Tumor volume (mm 3 )=(major axis) × (minor axis) × (height) ×0.52. (A) Effect of combined therapy with ISTF and anti-4-1BB mAb on Renca tumor volume. Each line indicates the tumor volume of the individual animal. Results shown are representative of three independent experiments. n, number of mice per experiment. (B) Representative images of tumors on days 21 in the same condition as (A). (C) Serum ALT and AST were measured. (D) Survival rate of mice was determined. The animals were sacrificed when the longest dimensions of the tumors were >20 mm. Comparison of survival curves with log-rank test yielded a statistical significance of ***P<0.001. NS, not significant; ISTF, immunostimulatory factor; mAb, monoclonal antibody; i.p., intraperitoneally; ALT, alanine aminotransferase; AST, aspartate aminotransferase.

Article Snippet: Renca cells were purchased from the Korean Cell Line Bank (Korean Cell Line Research Foundation) and cultured in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.).

Techniques: Control, Injection, Comparison

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Unveiling CXCR2 as a promising therapeutic target in renal cell carcinoma: exploring the immunotherapeutic paradigm shift through its inhibition by RCT001

doi: 10.1186/s13046-024-02984-2

Figure Lengend Snippet: RCT001 efficiently inhibits RCC growth and synergizes with ICIs. RENCA cells were injected subcutaneously. Once the tumor reached 30 mm 3 , mice were divided into different groups: Control ( n = 10), RCT001 alone ( n = 10, 20 mg/kg), anti-PD-1 + anti-CTLA4 ( n = 10, 100 μg each) and the combination of RCT001 + PD-1 + anti-CTLA4 ( n = 10). a Tumor volume was measured each day. Results are expressed as mean ± sd. b Tumor weights at the end of the experiment. c Blood vessels were visualized and quantified by IHC for αSMA d Tumor-associated macrophages (TAMs, total population), M2 TAMs and M1 TAMs were evaluated by cytometry. e The specific mRNA markers of M2 TAMs (CCL17 and CCL22) and mRNA CXCR2 were determined by qPCR. f Tumor-associated neutrophils (TANs, total population) and mature TANs were evaluated by cytometry. g Intratumor dendritic cells (DCs, total population) and activated dendritic cells were evaluated by flow cytometry. h Intratumor natural killer (NKs, total population) and activated natural killer were evaluated by cytometry. i Intratumor T cells (total population) and CD4 + T, Activated CD4 + T were evaluated by flow cytometry. j Intratumor activated T cell specific mRNA marker (INFγ) mRNA was evaluated by qPCR. k Intratumor CD8 T, activated CD8 T and anergic CD8 T were evaluated by flow cytometry. l Intratumor anergic T cell specific mRNA marker (PD-L1) mRNA was evaluated by qPCR. Statistics were performed using the ANOVA test: * p < 0.01, ** p < 0.01, *** p < 0.001

Article Snippet: A total of 200,000 RENCA cells were subcutaneously inoculated into the flanks of female BALB/c mice obtained from Janvier Labs.

Techniques: Injection, Control, Cytometry, Flow Cytometry, Marker