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Identification and enrichment of cell subpopulations

A technology of cell populations and cells, applied in the field of isolation or enrichment, identification of tumor-initiating cells, and characterization

Inactive Publication Date: 2013-09-18
ABBVIE STEMCENTRX LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Unfortunately, such techniques and associated methodologies rely heavily on the ability to efficiently and reliably identify, characterize, and / or compartmentalize, isolate, or enrich for subpopulations of tumor-perpetuating cells that can then be efficiently analyzed

Method used

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  • Identification and enrichment of cell subpopulations
  • Identification and enrichment of cell subpopulations
  • Identification and enrichment of cell subpopulations

Examples

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example 1

[0250] Characterization of tumor-initiating cell populations

[0251] To characterize the cellular heterogeneity of solid tumors as they exist in cancer patients, elucidate the identity of tumor perpetuating cells (TPCs; i.e. cancer stem cells: CSCs) using specific phenotypic markers and identify clinically relevant diagnostic biomarkers and therapeutic targets , using art-recognized techniques to develop and maintain large non-traditional xenografts (NTX TM ) tumor bank. NTX tumor repertoires containing large numbers of discrete primary tumors proliferate in immunocompromised mice through multiple passages of heterogeneous tumor cells originally derived from obtained by cancer patients. The availability of large numbers of discrete early passage NTX tumor cell lines with well-defined lineages has greatly facilitated the identification and isolation of TPCs into NTX tumor lines whose cells closely mirror the biology of tumors taken directly from patients, allowing reproducib...

example 2

[0258] Enrichment of tumor-initiating cell populations by FACS and transplantation

[0259] In tumors showing heterogeneous expression of specific proteins or proteins of interest (eg, CD46 and / or CD324), cells are enriched or isolated based on such markers and then transplanted into immunocompromised mice. More specifically, to determine whether high or low levels of surface CD46 and / or CD324 might be associated with enhanced tumorigenicity, NTX tumor samples were dissociated using prior art techniques as described above and analyzed using FACSAria flow cytometry. (BD Biosciences) to provide subpopulations enriched for different markers, which were then transplanted into immunocompromised mice. In this aspect, cells are injected subcutaneously into the mammary fat pad of immunocompromised female recipient NOD / SCID mice at doses typically ranging between 1,000 to 50 cells per mouse. When tumors arising from these grafts reach 800-2,000mm 3 , mice were euthanized and tumors w...

example 3

[0262] Representative cell surface markers expressed on tumor-initiating cells

[0263] Tumor-initiating cells, like all cells, can express thousands if not hundreds of markers or antigens, preferably on their cell surface. As such, while CD46 and CD324 are preferred markers for TICs, other concomitant markers can be used to identify and / or isolate the same subpopulation of tumor cells independent of the use of molecules capable of detecting CD46 and / or CD324. After confirming the TIC population using the cell surface marker profile described above, the PhenoPrint array was repeated using reagents in order to identify proteins that showed expression profiles comparable to CD46 and / or CD324, suggesting that these proteins could be used as CD46 and CD324 for purposes of the present invention. / or alternatives to CD324.

[0264] To identify proteins that are associated with TIC cell populations or that are abundantly co-expressed with CD46 and / or CD324, include non-PE-conjugate...

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Abstract

Markers useful for the identification, characterization and, optionally, the enrichment or isolation of tumorigenic cells or cell subpopulations are disclosed.

Description

[0001] cross-referenced application [0002] This application claims the benefit under 35 U.S.C.119(e) of U.S. Provisional Application No. 61 / 380,181, filed September 3, 2010, and U.S. Provisional Application No. 61 / 510,413, filed July 21, 2011, each of which It is hereby incorporated by reference in its entirety. field of invention [0003] The present invention generally relates to the identification, characterization, and optionally isolation or enrichment of cells or subpopulations of cells, especially tumor initiating cells derived from various tumors. In selected aspects, the invention also relates to methods of isolating tumor-initiating cells and the resulting preparations, and to the use of tumor-initiating cells and cell populations for pharmaceutical research and development and for diagnostic, therapeutic and other clinical and non-medical applications. Various methods in clinical application. Background of the invention [0004] Stem and progenitor cell differ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395C07K16/40G01N33/574C07K16/30C07K16/22A61K49/00
CPCC12N5/0693G01N33/57484G01N2333/705G01N33/5011
Inventor S.J.戴拉M.桑塔圭达W.C.安德森
Owner ABBVIE STEMCENTRX LLC
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