Method, kit, stem cells and application for preparing autologous hematopoietic stem cells

A technology of hematopoietic stem cells and somatic cells, applied in the field of biomedicine, can solve the problems of low safety, less available quantity, insufficient sources of hematopoietic stem cells, etc., and achieve the effect of production speed and safety advantages.

Active Publication Date: 2016-07-06
深圳百年干细胞技术研究院有限公司
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Aiming at the problems of insufficient sources of autologous hematopoietic stem cells in the prior art, few available quantities, and low safety, according to one aspect of the present invention, a method for preparing autologous hematopoietic stem cells is provided, the method comprising:

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method, kit, stem cells and application for preparing autologous hematopoietic stem cells
  • Method, kit, stem cells and application for preparing autologous hematopoietic stem cells
  • Method, kit, stem cells and application for preparing autologous hematopoietic stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Preparation of Example 1 Kit

[0060] Operate in a safe operation table with a cleanliness of 10-100, and prepare at a low temperature of 4-10 degrees. In 500ml of RPMI640 medium, add: 10μM Y-27632 (C14H21N3O 2HCl); 10ng / mL stem cell factor; 10ng / mL interleukin 3; 10ng / mL interleukin 6; 10ng / mL interleukin 11; 10ng / mL macrophage colony stimulating factor (M-CSF); 10ng / mL granulocyte colony stimulating factor (G-CSF); 10 μg / mL fucoidan; 10 μg / mL sulfuric acid Dextran; 10 nM AMD3100 (1,1′-[1,4-phenylenebis(methylene)]-di-1,4,8,11-tetraazacyclotetradecane); 10 μM M-alendronate sodium trihydrate; 10 μM disodium pamidronate.

[0061] Mix well, dissolve, and then filter and sterilize through a 0.22-micron pore size filter to prepare a cell culture solution.

[0062] The cell culture fluid is placed in the kit to prepare a kit for producing protein-induced autologous hematopoietic stem cells.

Embodiment 2

[0063] Example 2 Preparation of protein-induced autologous hematopoietic stem cells (M-PiHPS) in mice

[0064] 3-5 month old Balb / C male mice (purchased from School of Radiology, Chinese Academy of Medical Sciences (Tianjin), animal certificate number: SCXK (Tianjin) 2010-0002) were selected, and all their blood was collected and centrifuged using conventional Ficoll The technology centrifuges and separates blood to obtain blood mononuclear cells. The obtained mononuclear cells were divided into 5 × 10 6 The density of cells / ml was inoculated in a cell culture dish, and the cell culture solution prepared in Example 1 was added to it, at 37°C and 5% CO. 2 Incubator for 5 days to obtain protein-induced autologous hematopoietic stem cells. Gently pipetting the cultured cells to completely suspend the cells, collect the cells in a centrifuge tube, and perform the centrifugation-physiological saline suspension cell washing operation. Sexual autologous hematopoietic stem cells. ...

Embodiment 3

[0065] Example 3 Preparation of human protein-induced autologous hematopoietic stem cells (M-PiHPS)

[0066] 50ml of human peripheral blood was collected, and the blood was centrifuged and separated using conventional Ficoll centrifugation technology to obtain blood mononuclear cells. The obtained mononuclear cells were divided into 5 × 10 6 The density of cells / ml was inoculated in a cell culture dish, and the cell culture solution prepared in Example 1 was added to it, at 37°C and 5% CO. 2 Incubator for 5 days to obtain protein-induced autologous hematopoietic stem cells. Gently pipetting the cultured cells to completely suspend the cells, collect the cells in a centrifuge tube, and perform the centrifugation-physiological saline suspension cell washing operation. Sexual autologous hematopoietic stem cells. The obtained protein-induced autologous hematopoietic stem cells were 1 × 10 5 pcs / ml to 1×10 6 The cell density of cells / ml was mixed with an equal volume of a mixt...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a method for preparing autologous hematopoietic stem cells. The method includes following steps of cultivating somatic cells in a cell culture fluid for 3-6 days and collecting the generated autologous hematopoietic stem cells. The cell culture fluid is an RPMI1640 culture fluid containing: 1-100 [mu]M of Y-27632 (C14H21N3O.2HCl), 1-100 ng / ml of stem cell factors, 1-100 ng / ml of interleukin 3, 1-100 ng / ml of interleukin 6, 1-100 ng / ml of interleukin 11, 1-100 ng / ml of a macrophage colony stimulating factor (M-CSF), 1-100 ng / ml of a granulocyte colony-stimulating factor (G-CSF), 1-100 [mu]g / ml of fucoidin, 1-100 [mu]g / ml of dextran sulfate, 1-100 nM of AMD3100 (1,1'-[1,4-phenylenebis(methylene)]-bis-1,4,8,11-tetrazacyclotetradecane), 1-100 [mu]M of M-alendronate sodium (Malendronate sodium trihydrate), and 1-100 [mu]M of disodium pamidronate. The invention provides a method for preparing the autologous hematopoietic stem cells, a kit, the stem cells and an application. In addition, the method is advantaged in yield, purity, and producing speed of the production of the protein-induced autologous hematopoietic stem cells.

Description

technical field [0001] The present invention relates to the field of biomedicine, in particular, to a method, kit, stem cell and application for preparing autologous hematopoietic stem cells. Background technique [0002] Stem cells are a kind of pluripotent cells with self-replication ability, which can differentiate into a variety of functional cells under certain conditions. According to the developmental stage of stem cells, they are divided into embryonic stem cells and adult stem cells. Embryonic stem cells are a class of early embryonic cells with unlimited self-renewal and differentiation potential. They are cells with inner cell patterns when the fertilized egg divides and develops into blastocysts. They have the ability to differentiate into cells in various germ layers. Adult cells refer to stem cells with repair and regeneration ability that exist in specific tissues or organs of humans and mammals (including bone marrow, nerve, muscle, epidermis, testis or kidn...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0789C12N5/074A01N1/02A23L33/00A61K35/28A61P39/06A61L27/38
Inventor 林雄斌林柳吟
Owner 深圳百年干细胞技术研究院有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap