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In Vivo Cryopreservation Technique Detects Human Tumor Tissues in Xenograft Mouse Models

SHIRLEY, NEW YORK (NY), USA, July 20, 2015 / -- As early as in 1889, Stephen Paget proposed the "seed and soil" hypothesis which laid a foundation for the concept of tumor microenvironment. As an extension of the "seed and soil" hypothesis, Paget predicted accurately that if the "seed" tumor cell can settle in the "soil" for the growth, namely remote organ, tumor cells must synergistically impact with the around factors.

Micro environment is a complicated system. It consists of a number of stromal cells, including the fiber cells, immune and inflammatory cells, fat cells, glial cells, smooth muscle cells and vascular cells. These cells can be induced by tumor cells to produce a large number of growth factors, cytokines and matrix degrading enzymes, which are beneficial to the growth and invasion of tumor cells.

In this study, we used IVCT and Cb for immunohistochemical analysis of soluble plasma protein on human cancer cell xenograft mouse models, and compared with other traditional preparation methods, such as FQF and IMDH. In addition, we also used bovine serum albumin (BSA) tail vein injection to observe the dynamic distribution of soluble protein in tumor tissues prepared by IVCT.

Objective: the mechanism through the stroma transfer of human tumor soluble molecules is related to in vivo tumor behavior closely. But in the traditional organization preparation, artifact formed in the process hinders the detection of soluble components in tissue. In the present study, "in vivo cryotechnique" (IVCT) and "frozen biopsy" (CB), these two methods are used respectively in the xenograft human tumor cell models. Immune segment of exogenous and endogenous plasma components are detected that the two techniques can make the target tissue in vivo cryofixation.

Methods: after injecting the human lung cancer cells subcutaneously into the nude mice, using different methods to prepare tumor tissue paraffin sections and frozen sections and detect plasma proteins with immunolocalization, such as albumin, immunoglobulin G1 and immunoglobulin M, and bovine serum albumin (BSA) via the tail vein injection.

Results: compared with the traditional method, IVCT and Cb can more clearly observe the stroma of the tumor cells and extracellular matrix. In addition, the bovine serum albumin via tail vein injection can ooze from capillary leakage to the connective tissue around the tumor within 24 hours and spread to the tumor stroma and extracellular matrix after 3 days gradual injection.

Conclusion: the protein molecules from the capillary leakage in the progression of tumors in different parts changed greatly and small molecular plasma proteins (non large molecular proteins) massively distributed in the tumor extracellular matrix. Study on the immunohistochemical study of soluble proteins in tumor blood circulation shows that IVCT and Cb are more favorable for the analysis of the tissue pathology than the traditional technology.

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Andrea Brook
Creative Animodel
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