Journal: Cancers

Article Title: Therapeutically Engineering Exosomes to Target CD206+ M2 Macrophages to Prevent the Development of Primary Tumors and Distal Metastases in Breast Cancers

doi: 10.3390/cancers18101619

Figure Lengend Snippet: Differential biodistribution of exosomes and protein in 4T1 tumors (Balb/c mice): I-131-labeled control exosomes (HEK293 exo), anti-mouse CD206 antibody (CD206 Ab), fusion protein (fusion P) containing targeting peptide and Fc-mIgG2b, and engineered exosomes (Engi. Exo) carrying targeting peptide were administered IV into 4T1 breast cancer-bearing animals; SPECT images were obtained 3 h after administration. ( A ) SPECT images showed the distribution of the agents in the primary tumors (white circles) and lungs (yellow rectangular areas). White arrows show possible metastatic foci. Fusion protein and engineered exosomes accumulated in the primary and metastatic tumors. We used rainbow color to depict the activity of I-131. Blue is the lowest and red is the highest activity of I-131. ( B ) Randomly selected lung H&E staining showed multiple metastatic foci (black arrows). ( C ) Presence of numerous CD206+ cells is seen in tumors (green cells). n = 2–3 animals per group. Ab = antibody, Engi = engineered, Exo = exosomes, HE = Hematoxylin and Eosin, P = protein, SPECT = single-photon emission computed tomography, H = heart.

Article Snippet: Syngeneic, aggressive triple-negative breast cancer (TNBC) mouse cell lines 4T1 (for Balb/c mice, ATCC, Manassas, VA, USA) and AT3 (for C57BL/6, Millipore, Burlington, MA, USA) were obtained and maintained in the laboratory in Dulbecco’s Modified Eagle Medium (DMEM) (Corning, Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS) (Nalgene-GIBCO, Rochester, NY, USA), sodium pyruvate (GIBCO, Grand Island, NY, USA), non-essential amino acids (GIBCO), sodium glutamate (GIBCO), and 100 U/mL penicillin and streptomycin (GIBCO).

Techniques: Labeling, Control, Single Photon Emission Computed Tomography, Activity Assay, Staining

Journal: Cancers

Article Title: Therapeutically Engineering Exosomes to Target CD206+ M2 Macrophages to Prevent the Development of Primary Tumors and Distal Metastases in Breast Cancers

doi: 10.3390/cancers18101619

Figure Lengend Snippet: Resected model of breast cancer (4T1 in Balb/c mice) and metastasis: Representative images from tumor-resected animals (two from each group). All the primary tumors were resected 11 days after tumor cell implantation, and the animals were followed up to 63 days after resection. The figure shows representative animals from the control (HEK293 ( left panel )) and engineered exosome-treated ( right panel ) groups. The animals treated with the control exosomes began showing local recurrence and lung metastasis (black arrows show local recurrence and multiple distal metastases) starting on day 28 post-resection. None of the animals that received the engineered exosomes to deplete CD206+ cells showed any recurrence or distal metastasis. It is noteworthy that a small leftover tumor (white arrow) in one of the engineered exosome-treated animals disappeared following therapy on subsequent bioluminescent imaging. n = 5 per group. Pre-Sur = pre surgery, POD = post-operative day.

Article Snippet: Syngeneic, aggressive triple-negative breast cancer (TNBC) mouse cell lines 4T1 (for Balb/c mice, ATCC, Manassas, VA, USA) and AT3 (for C57BL/6, Millipore, Burlington, MA, USA) were obtained and maintained in the laboratory in Dulbecco’s Modified Eagle Medium (DMEM) (Corning, Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS) (Nalgene-GIBCO, Rochester, NY, USA), sodium pyruvate (GIBCO, Grand Island, NY, USA), non-essential amino acids (GIBCO), sodium glutamate (GIBCO), and 100 U/mL penicillin and streptomycin (GIBCO).

Techniques: Control, Imaging

Journal: Animal Models and Experimental Medicine

Article Title: Characterizing a murine breast cancer mouse model reveals chromosomal abnormalities in structure and number of single‐cell clones and the presence of rare cancer stem cell‐like phenotypes

doi: 10.1002/ame2.70156

Figure Lengend Snippet: A 4T1 BALB/c mouse model of breast cancer. The left flanks of mice were injected with 4T1 cells subcutaneously. (A) A representative picture of an injected mouse is shown with a white arrow indicating the site of primary tumor growth. (B) The tumor volume was evaluated twice per week. (C) A representative excised tumor. (D) H&E staining of primary tumor tissue at the end point of tumor development with pleomorphic vesicular nuclei (green arrows) and hyperchrome prominent nucleoli (yellow arrows) indicated (40× magnification, and the scale bar represents 20 μm).

Article Snippet: The 4T1 mouse breast cancer cell line was purchased from the American Type Culture Collection (ATCC R CRL‐2539) (Manassas, Virginia, USA).

Techniques: Injection, Staining

Journal: Animal Models and Experimental Medicine

Article Title: Characterizing a murine breast cancer mouse model reveals chromosomal abnormalities in structure and number of single‐cell clones and the presence of rare cancer stem cell‐like phenotypes

doi: 10.1002/ame2.70156

Figure Lengend Snippet: Chromosome counts of metaphase cells. (A) Chromosome number of murine 4T1 breast cancer cells prepared ex vivo (23 cells) and in vitro (100 cells). Boxes represent data range from 1st quartile to 3rd quartile. Whiskers represent the range of chromosomes from minimum to the maximum. (B) Descriptive statistics and Mann–Whitney two‐tailed test results. (C) Distribution of chromosome numbers in ex vivo and in vitro cells.

Article Snippet: The 4T1 mouse breast cancer cell line was purchased from the American Type Culture Collection (ATCC R CRL‐2539) (Manassas, Virginia, USA).

Techniques: Ex Vivo, In Vitro, MANN-WHITNEY, Two Tailed Test

Journal: Animal Models and Experimental Medicine

Article Title: Characterizing a murine breast cancer mouse model reveals chromosomal abnormalities in structure and number of single‐cell clones and the presence of rare cancer stem cell‐like phenotypes

doi: 10.1002/ame2.70156

Figure Lengend Snippet: Analysis of breast cancer stem cell‐like phenotypes (CD44 + /CD24 − ). (A) Flow cytometric analysis of CD24/CD44 expression in ex vivo and in vitro cells. (B) Percentage of four subpopulations in breast cancer ex vivo and in vitro 4T1 cells. Standard deviation is indicated for each respective cell suspension, n = 3.

Article Snippet: The 4T1 mouse breast cancer cell line was purchased from the American Type Culture Collection (ATCC R CRL‐2539) (Manassas, Virginia, USA).

Techniques: Expressing, Ex Vivo, In Vitro, Standard Deviation, Suspension