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Human osteosarcoma stem-cell related antigenic marker and application thereof

An osteosarcoma and marker technology, applied in the field of tumor biology, can solve the problems of high instrument configuration, dye toxicity, low cell viability, etc.

Active Publication Date: 2011-05-11
RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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Problems solved by technology

On the one hand, the work of the above-mentioned scholars is mainly limited to the level of in vitro cell culture, failing to establish an osteosarcoma cell xenograft model in vivo to identify the phenotype and function of osteosarcoma stem cells (which is the gold standard for cancer stem cell research); On the other hand, the separation method of SP cells has limitations such as hoechst33342 dye toxicity, low cell viability, and high instrument configuration.

Method used

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  • Human osteosarcoma stem-cell related antigenic marker and application thereof
  • Human osteosarcoma stem-cell related antigenic marker and application thereof
  • Human osteosarcoma stem-cell related antigenic marker and application thereof

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Embodiment Construction

[0015] I. In our previous study, we used the primary specimens of primary osteosarcoma patients, digested them into single cell suspensions, and successfully established osteosarcoma cell spheres and cross-species using serum-free culture technology and NOD / SCID mice. Transplant the model. Phenotype analysis of osteosarcoma cells from tumor-forming mice was performed by flow cytometry, and it was found that these cells often expressed SSEA-3, SSEA-4, CD184, CD105, CD166 and CD49d, etc. Stem cells and embryonic stem cell surface markers.

[0016] II. More detailed verification of SSEA-4: using cell lines, using immunofluorescence and immunohistochemical methods, for phenotypic identification and verification, and testing the expression in primary specimens, etc.

[0017] III. Through a more detailed distinction and comparison between different surface markers, select a small part of the most specific markers for immunofluorescence antibody labeling, and use flow sorting techno...

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Abstract

The invention discloses a human osteosarcoma stem-cell related antigenic marker stage-specific embryonic antigen-4 (SSEA-4) and an application thereof in a targeted therapy of osteosarcoma. The SSEA-4 is a specific osteosarcoma stem-cell surface marker which is almost not expressed in each normal adult tissue. The surface marker is easy for antibody labeling without affecting the cell viability, and applicable to the function tests after separation, therefore, a reliable molecular tracer is provided for further researching the function of stem cells in the origin, occurrence and development processes of osteosarcoma, and a foundation for explaining the clinical osteosarcoma drug resistance as well as transferring and looking for cellular and molecular targets for the targeted therapy is laid.

Description

technical field [0001] The present invention relates to the field of tumor biology, more specifically, relates to a human osteosarcoma stem cell-associated antigen marker and application thereof. Background technique [0002] In recent years, with the deepening understanding of the theory of tumor stem cells, we are required to have a clearer understanding of the origin, occurrence and development of tumors. In particular, reports on the isolation and identification of tumor stem cells such as leukemia, breast cancer, human glioma, and colorectal cancer not only confirmed and consolidated the theory of tumor stem cells, but also enabled the identification of other tumor stem cells and the study of the characteristics of their internal molecular mechanisms. become hot. At present, there are very few reports on osteosarcoma stem cells at home and abroad. In 2005, Gibbs et al. used in vitro culture to isolate sarcospheres containing stem cell-like osteosarcoma cells from cell ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/532G01N33/533
Inventor 张武诸江张伟宾
Owner RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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