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Compositions enriched in neoplastic stem cells and methods comprising same

A technology for stem cells and compositions, applied to compositions rich in neoplastic stem cells and fields containing the same, can solve problems such as troubles, damage to cell viability, inability to detect pure cell populations, and the like

Inactive Publication Date: 2009-08-05
UNIV OF TENNESSEE RES FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Other methods of cell identification and subsequent isolation and / or enrichment such as gel electrophoresis cannot probe pure cell populations, are plagued by contamination and / or impair cell viability

Method used

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  • Compositions enriched in neoplastic stem cells and methods comprising same
  • Compositions enriched in neoplastic stem cells and methods comprising same
  • Compositions enriched in neoplastic stem cells and methods comprising same

Examples

Experimental program
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Effect test

Embodiment 1

[0137] Frequency of OCT4-expressing cells in tumors

[0138] To better investigate the frequency of OCT4-expressing cells in the tumors, immunohistochemical analysis was performed on serial sections derived from the following tumors: osteosarcoma, glioblastoma and ductal carcinoma. figure 1The results shown indicate that OCT4-positive nuclei are present in osteosarcoma and glioblastoma; additionally, Image 6 The results shown in A-B indicate that OCT4-positive nuclei are also present in ductal carcinoma and breast cancer metastases to the brain, respectively. Although OCT4-positive cells were observed in both ductal carcinoma and breast cancer metastases, compared with primary breast cancer confined to ductal carcinoma ( Image 6 A), OCT4-positive nuclei appear more frequently in breast cancer metastases to the brain ( Image 6 B). Both tumors and metastases contain OCT4-positive cells, thus the methods described herein demonstrate that these OCT4-positive cells are eff...

Embodiment 2

[0140] OCT4, Nanog, and STAT3 are consistently expressed in glioblastoma clinical samples and cell lines

[0141] To determine whether OCT4, Nanog and STAT3 were co-expressed in glioblastoma clinical samples and in the cell lines LN18, LN229, LN428 and U251, semi-quantitative RT-PCR analysis followed by western blot analysis was performed. figure 2 A shows results showing moderate to high level mRNA expression of OCT4, Nanog and STAT3. The expression level of the protein is the same as that of figure 2 B shows the mRNA expression levels associated with, figure 2 Moderate to high protein expression levels of OCT4, Nanog and STAT3 are demonstrated in B.

Embodiment 3

[0143] OCT4 expression in stromal-attached or unattached growing tumor cells

[0144] To examine the effect of cell-matrix attachment on OCT4 expression in tumor cells, osteosarcoma and mammary tumor cells were grown as sarcospheres or mammospheres in matrix-attached or non-attached form, respectively. image 3 The results shown indicate that hi OCT4 and Nanog expression is dependent on cell attachment, thus tumor cells growing unattached in the form of sarcospheres and mammospheres express OCT4 and Nanog at high levels, in contrast to their inhibitory effects in stromal-attached tumor cells, in The expression of Nanog and OCT4 was relatively low in the latter case.

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Abstract

A neoplastic stem cell population enriched for expression of the OCT4 transcription factor as well as methods for their identification, isolation and enrichment are described. The OCT4-enriched neoplastic stem cell population is further utilized for the induction and analysis of cancer in an animal. In addition, methods of preventing, abrogating, or inhibiting cancer, tumor growth, and metastasis via OCT4 inhibition are further provided.

Description

Background of the invention [0001] To date, methods for the analysis of neoplastic cells are neither efficient nor uniform enough for research purposes. Neoplastic stem cells isolated from tissue samples by various fractionation procedures consist of mixed types of cells. Efficient isolation of neoplastic stem cells can provide the means to explore fundamental mechanisms of cancer cell biology and disease. There is a great need in the art for methods to specifically and efficiently isolate and proliferate subpopulations of cells to provide large neoplastic stem cell populations for in vitro and in vivo studies. [0002] Identification of stem cell markers in neoplastic cells provides valuable information for a variety of clinical and basic research applications. Identification and subsequent isolation of neoplastic stem cells (NSCs) from specific tumors or metastases can be used, for example, to diagnose disease and / or develop rational therapeutic measures for developing con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12N5/09C12N5/095
CPCC12N5/0695C12N2501/115C12N2502/1323C12N2501/33C12N2501/11C12N2503/02C12N5/0693G01N2333/4703G01N33/574C12N2503/00A61P35/00A61P35/04A61P37/00
Inventor C·丁奇V·屈克考夫T·伊甘托娃
Owner UNIV OF TENNESSEE RES FOUND
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