Multicolor flow cytometry method for identifying population of cells, in particular mesenchymal stem cells

A technology of cell populations and mesenchymal stem cells, applied in animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve problems such as inflexibility and inability to apply to individual research requirements, and achieve the effect of improving detection sensitivity

Inactive Publication Date: 2016-03-02
CELL THERAPUETICS INC
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Problems solved by technology

[0004] Published examples of multicolor reagent panels for MSC to date ignore recommended ISCT criteria for cell markers (Martins et al., 2009; Jones et al., 2006), utilize parallel tubes rather than single-tube approaches (Tucker and Bunnell, 2011) or utilize custom-made conjugates and the most common fluorochromes (Bradford and Clarke, 2011) resulting in an inflexible tool that cannot be adapted to individual research requirements

Method used

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  • Multicolor flow cytometry method for identifying population of cells, in particular mesenchymal stem cells
  • Multicolor flow cytometry method for identifying population of cells, in particular mesenchymal stem cells
  • Multicolor flow cytometry method for identifying population of cells, in particular mesenchymal stem cells

Examples

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Embodiment

[0095] Materials and methods

[0096] Collection of Human Umbilical Cord Matrix Samples

[0097] Human umbilical cords and placentas were collected from term births following elective cesarean delivery and stored aseptically at room temperature for less than 90 minutes during transport (from delivery to handling). The umbilical cord was separated from the placenta, and the 10cm section near the placenta was removed and placed in a sterile container. Umbilical cords were washed with phosphate buffered saline (PBS; Life Technologies Ltd, Paisley, UK) to wash away blood and incubated at 4°C in Hanks buffered saline (HBSS) supplemented with antibiotic-antimycotics (both from Life Technologies) 2 hours. The study was approved by the local research ethics committee, and all mothers gave written informed consent. Umbilical cord stromal MSCs were prepared according to published methods with partial modifications [13,14]. Then the cells were collected and counted, and the cells o...

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Abstract

This invention is in the field of the identification and even isolation of mesenchymal stem cells (MSCs) and other cell types by means of differential specific fluorescence activated cell sorting (FACS).

Description

technical field [0001] The present invention is in the field of identification and subsequent isolation of mesenchymal stem cells (MSCs) and other cell types by means of differentially specific fluorescence activated cell sorting (FACS). That is, the present invention is an agent or composition of matter, at various relative specific concentrations, that unexpectedly provides the labeled cells with specific concentrations of the labeled cell marker in the reagent. Medical (or Physiological) Effects. The present invention is reagents and methods for the selective isolation of cells by means of specific relative reagent concentrations. The present inventors have provided a surprising means by which a composition of matter can be made to act differentially on cells as a drug, thereby enabling the use of fluorescence-activated cell sorting (FACS) to identify more than seven types of cell surface Cells are identified simultaneously in the presence or absence of markers. Backgro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775G01N33/50
CPCC12N5/0663C12N5/0665G01N2333/70596G01N33/533G01N33/56966G01N2333/705
Inventor M·J·埃文斯A·雷金纳德S·苏丹
Owner CELL THERAPUETICS INC
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