Kineto Lab services

Kineto Lab Ltd. is an acknowledged expert in development of tailored, predictive screening procedures employing primary patient tumors, xenograft assays and cell based preclinical screening procedures for anticancer agents.

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In vitro and in vivo technical facilities and capabilities of Kineto Lab cover the successful investigation of all the steps of the metastasis process:

1/ In-vivo and in-vitro models of metastasis

• Spontaneous models of metastasis: tumor cells have to complete all steps of the metastatic process

•  Site of injection (target organs): subcutaneous tissue (lung), spleen (liver), hind leg muscle (lung). 

• Orthotopic transplantation of tumor cells (target organs): wall of large bowel (liver), kidney (lung), mammary   fat pad (lung), dermis (lung and/or lymphnodes).

• Experimental models of metastasis: Site of injection: tail vein, portal system, carotid artery, left ventricle of the heart. Target organs are: lung, liver, brain, all organs, respectively.

• In-vitro models of the steps of the metastatic process.

• Tumor cell lines used: human and mouse melanomas, fibrosarcomas, colon carcinomas, lung carcinomas, breast carcinomas, prostate carcinomas, lymphomas, HNSCC.

2/ Tissue invasion

2D models:

• confluent cell cultures of normal cells (fibroblasts, hepatocytes, different epithelial cells) confronted with tumor cell of different origin.

• adhesion of tumor cells to cryosections from different organs

3D models:

• organ cultures/regenerated organ fragments (chorioallantoic membrane, diaphragm, human amnion, embryonic skin, lung cubes.

• Intravasation: isolated brain capillaries are confronted with different tumor cell lines.

• Extravasation: interaction of cultures of endothelial cells or isolated vessel fragments with tumor cells.

3/ Analysis of tumor cell motility

• Types of cellular motility: chemotaxis (liquid phase gradient), haptotaxis (solid phase gradient), chemokinesis (random motility).

Migration tests:

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• 2D: cell culture wounding, phagokinetic track assay, video microscopy, emigration of cells from tumor spheroids (scatter).

• 3D: Boyden chamber (cell number is determined on the lower side of the 8 μm pore size membrane), migration into extracellular matrix gels (collagen I, fibrin, basement membrane), organ cultures (chorio-allantoic membrane, denuded cornea).

4/ Assessment of anti-angiogenic potential.

• Frozen tumor samples growing at different location are cryosectioned. Immunflorescence staining is performed to highlight basement membrane and CD31 to unequivocally detect vessels. Vessel density (blood, lymphatic) vessel size, area fraction are determined using image analysis systems.

5/ Assessment of anti-metastatic potential.

• Metastases of different tumor lines are produced in spleen-liver, muscle-lung, skin-lung models. The removed target organs are fixed and number and size of the surface colonies are determined. If necessary, the target organs are serially sectioned and the area fraction of the metastatic colonies is determined.

6/ Detection of proliferation and apoptosis

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• Processing and evaluation of Pathology Samples

• Pathology sample processing

• Microscpic analysis of frozen sections

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Cell Lines

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Metastasis model systems and orthotopic transplantations (samples)

I. Brain metastases

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1. Macroscopic meatastases are produced by injecting tumor cells directly in the brain parenchima

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2. Microscopic metastases are produced by injecting tumor cell in the carotid artery

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II:  Pleural metastases can be produced by injecting tumor cell in the thoracic cavity. Metastases can be analyzed by dissecting the diaphragm

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III. Colon carcinoma: Tumor pieces are transplanted onto the

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IV. Mammary carcinoma (injected into mammary fat pad)

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V. Prostate carcinoma (tumor cells are injected into the

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VI. General metastasis model:

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Tumor cells are injected into the left ventricle of the heart.

Metastases are formed in all organs.

VII. Colonization models:

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Tumor cells are injected into the tail vein, or spleen, and tumor colonies are formed in lung and liver, respectively.

VIIa. Amputation metastasis model which is used to accelerate metastasis formation:

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Tumor cells are injected into the foot pad. The injected leg is amputated after primary tumor is formed. Metastases develop in the lungs.