Establishment and Characterization of a New Canine Mast Cell Tumor Cell Line.

Abstract

ABSTRACT. A new cell line (CoMS) was established from a 3-year-old male mongrel dog with mast cell tumor of the oral mucosa. CoMS cells grow in suspension with a doubling time of 27.0 ± 0.7 hr. The cytoplasmic granules were formalin-sensitive, showed diverse appearances in their ultrastructural findings and contained heparin proteoglycan and neutral protease chymase. Calcium ionophore A23187, substance P and concanavalin A caused significant histamine release from CoMS cells, while compound 48/80 failed to release histamine. This cell line will make an available source for studies on canine mast cell tumors. KEY WORDS: canine, cell line, mast cell tumor.J. Vet. Med. Sci. 63(9): 1031-1034, 2001 Mast cell tumor (MCT) is one of the common neoplasms in dogs, accounting for 7% to 21% of all skin tumors and 11% to 27% of all malignant skin tumors [19]. There are still some dilemmas caused by its unpredictable biological behaviors with regard to diagnosis and treatment of MCT [9]. Studies on several neoplastic cell lines of human or rodent mast cells such as HMC-1, P-815 and RBL-2H3 have provided many informations about mast cell biology [17,18,20]. There will be anatomical, biochemical, immunological and pharmacological differences between human, rodent and canine mast cells. Available cell lines of canine MCT are therefore required for the research on this disease.In this paper, we describe a successful establishment of a new canine MCT cell line named CoMS in continuous culture and report morphology, histamine release activity and granular components such as proteases and heparin of CoMS cells.CoMS cell line was obtained from a mucosal mass in the lower lip of a 3-year-old male mongrel dog with MCT. The dog was referred to the Veterinary Teaching Hospital of Hokkaido University with massive swelling of a left lower lip and both cervical lymph nodes. The clinical managements were consisted of surgical excision, radiotherapy and chemotherapy. In spite of these treatments, the dog died of deterioration of general conditions 56 days after the first admission. Multiple gastroduodenal ulcers and multiple distant metastases to the internal organs such as the liver, spleen, kidneys and lungs but no metastasis to the skin were observed at necropsy.The specimen of CoMS collected at surgery was initially maintained in vivo passage in 5-week-old female C.B-17 severe combined immunodeficiency (SCID) mice (Clea Lab., Tokyo, Japan) by subcutaneous implantation, then the tumor tissues excised from the mice were used for establishment of the cell line in vitro.These tumor tissues were minced finely in RPMI 1640 medium containing 2 mg/ml sodium bicarbonate, 25 mM Hepes buffer, 100 U/ml penicillin and 100 µg/ml streptomycin, filtrated through a stainless mesh and washed two times in phosphate buffered saline. The isolated cells were then suspended and incubated in the above RPMI 1640 medium with 10% heat inactivated fetal calf serum (FCS) at 37°C in a humidified atmosphere of 5% CO 2 and were passaged every 4 to 6 day. The cell line n...

Expression of the MDR1 Gene and P-Glycoprotein in Canine Mast Cell Tumor Cell Lines

“…Frequency and intensity of the chemotherapy may potentially give influences on MDR1 expression, however, none of these factors correlated with MDR1 status of the cell lines investigated in this study [10,23]. Alternatively, TiMC cells may not show induction of MDR1 gene expression in response to some factors, similar to the previously reported MDR1-deficient genotype [13,20].…”

“…Cell lines: TiMC, CoMS and LuMC canine MCT cell lines were originated from a cutaneous MCT, an oralmucosal MCT and a gastrointestinal MCT, respectively [10,23]. CoMS cell line was kindly provided by Laboratory of Veterinary Surgery, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University.…”

Retinoids Induce Growth Inhibition and Apoptosis in Mast Cell Tumor Cell Lines

“…Cell culture: Three canine MCT cell lines (CoMS, CM-MC and VI-MC) were used in this study [9,21]. CoMS and VI-MC cells were cultured in RPMI-1640 medium (Nissui Pharmaceutical Co., Tokyo, Japan) supplemented with 10% heat-inactivated (56°C, 30 min) fetal bovine serum (FBS), 2 mM L-glutamine, 50 mg/L gentamycin sulfate and 1.5 mg/ L amphotericin B. CM-MC cells were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated FBS, 2 mM L-glutamine, 10 U/ml penicillin G, 10 µg/ml streptomycin and 50 mM 2-mercaptoethanol.…”

Molecular Cloning of Canine Membrane-Anchored Inhibitor of Matrix Metalloproteinase, RECK

“…Quantitative RT-PCR analysis of canine RECK gene expression in normal tissues and canine tumor cells: Reverse-transcription was performed using total RNA samples isolated from various tissues (brain, heart, lung, liver, spleen, small intestine, pancreas, kidney, skeletal muscle, and testis) of three adult healthy male beagles and five canine tumor cell lines: osteosarcoma [11], high metastatic osteosarcoma [3], transitional cell carcinoma, mammary gland tumor (our unpublished cell lines), and mast cell tumor [9], and RECK expression was analyzed by quantitative RT-PCR method.…”