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High purity mesenchymal stem cells

A technology of stem cells and cells, applied in the field of high-purity cell groups, can solve the problems of fluctuation of differentiation ability and proliferation ability, and achieve the effect of excellent differentiation ability

Pending Publication Date: 2022-08-02
PUREC CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] However, fluctuations in the differentiation ability and proliferation ability of REC clones can still be observed
In addition, even for RECs with excellent proliferative properties, deterioration due to passage cannot be avoided, and effective quality control methods are required
In addition, when the expression of Ror2 is used as an index, there is still room for improvement in the correlation with proliferation ability

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] 1. Determination of CV value of forward scattered light (FSC) based on flow cytometry

[0103] by known methods (WO2016 / 17795) and figure 1 In the indicated scheme, pre-prepared REC clones (clone numbers: 1 to 45) were used for flow cytometry-based forward scattered light CV value determination.

[0104] FSC in flow cytometry is proportional to the surface area or size of the cells. In this example, fluctuations in cell size were evaluated using the CV value of FSC as an index. exist figure 2 The results of the measurement of the CV value of the forward scattered light by flow cytometry are shown in FIG.

[0105](a) PI staining was performed on each REC clone, the gate was set to the PI-negative live cell population, and the dead cells were excluded from the analysis target.

[0106] (b) The PI-negative live cell population is expanded in the cell histogram of FSC / SSC, the target (P1) is set to the main cell population, and garbage and noise are excluded from the a...

Embodiment 2

[0123] Two bone marrow fluid batches (#18TL158166 and #8F5040) were sorted by two methods (CD271 and CD90 double positive and CD271 alone), and the resulting ratios of Colony, REC, MEC, SEC were compared ( Image 6 ).

[0124] exist Image 6 , the left panel represents the results of cell sorting. Cells from the framed portion were seeded into 96-well plates, and the resulting colony ratios are shown in "Colony" in the table to the right. Among the obtained colonies, the ratios of the colonies with rapid proliferation (REC), the colonies with moderate degree of proliferation (MEC), and the colonies with slow proliferation (SEC) are shown in the table.

[0125] Compared with the case of sorting with LNGFR and CD90 double-positive as indicators, when sorting with LNGFR R alone, the obtained community ratio was higher, but the obtained REC ratio was lower.

[0126] However, it was found that RECs can be isolated even when LNGFR positive alone is sorted.

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Abstract

A cell group which is a cell group of a highly proliferative mesenchymal stem cell clone in which LNGFR (CD271) is positive or LNGFR (CD271) and Thy-1 (CD90) are both positive, said cell group satisfying at least one of the following characteristics (a) and (b). (a) The coefficient of variation of forward scattered light in flow cytometry is 35% or less. (b) The average cell size is 20 [mu] m or less.

Description

technical field [0001] The present invention relates to a high-purity cell population of human mesenchymal stem cells with homogeneity and rapid proliferation. Background technique [0002] Mesenchymal Stem Cells (MSCs) have few ethical issues associated with cell collection, and have diverse differentiation capabilities into bone, cartilage, and fat, so they are somatic stem cells that have been widely used in clinical applications after hematopoietic stem cells. a kind of. Since MSCs can be isolated by a relatively simple method, they are mainly induced to differentiate into cartilage, bone, etc. in a test tube, and then transplanted locally, etc., and are widely used as a material for biomaterials. [0003] In addition, in advancing clinical applications, it is a necessary condition for commercialization to be able to manufacture cells that maintain a constant function in the required amount. [0004] However, when there are fluctuations in the properties of the startin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0663C12N5/0662C12N2330/10C12N2500/02C12N2501/115
Inventor 松崎有未陶山隆史
Owner PUREC CO LTD
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