Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Agent containing VDUP1 protein or its encoding gene for differentiating hemopoietic stem cell into natural killer cell, and method for differentiating hemopoietic stem cell into natural killer cell by

A technology for hematopoietic stem cells and coding genes, which is applied to a reagent containing VDUP1 protein or its coding gene for differentiating hematopoietic stem cells into natural killer cells, which can solve problems such as unknown effects.

Inactive Publication Date: 2008-06-11
医疗细胞公司
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

VDUP1 expression is dominant in immune cells, but its full role in immune cells is unknown

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
  • Agent containing VDUP1 protein or its encoding gene for differentiating hemopoietic stem cell into natural killer cell, and method for differentiating hemopoietic stem cell into natural killer cell by
  • Agent containing VDUP1 protein or its encoding gene for differentiating hemopoietic stem cell into natural killer cell, and method for differentiating hemopoietic stem cell into natural killer cell by
  • Agent containing VDUP1 protein or its encoding gene for differentiating hemopoietic stem cell into natural killer cell, and method for differentiating hemopoietic stem cell into natural killer cell by

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1: Isolation of hematopoietic stem cells from bone marrow

[0067] Bone including the tibia and femur of C57BL / 6 mice (Daehan Biolink, Korea) of 6-9 weeks was ground, and the ground product was passed through a 70 μm cell strainer, and a dissolving solution (Sigma, St.Louse , MO) to remove red blood cells to obtain bone marrow cells. Bone marrow cells were labeled with biotin-specific system markers (CD11b: macrophage marker, Gr-1: granulocyte marker, B220: B cell marker, NK1.1: NK cell marker, CD2: T cell marker, TER- 119: erythrocyte marker) antibody response, however, wash. The cells were reacted with streptavidin-labeled magnetic beads (Miltenyi Biotec, Auburn, CA). For magnetically labeled Lin + Cells were harvested by passing them through a CS column (Miltenyi Biotec) within the magnetic field of a SuperMACS (Miltenyi Biotec, Auburn, CA). Lin passing through the column - Cells react with magnetic beads coupled to c-kit. After passing through the MS c...

Embodiment 2

[0068] Example 2: Inducing HSCs to differentiate into NK cells

[0069] The HSCs isolated from bone marrow in the above-mentioned Example 1 were inoculated in 6-well plates (Falcon, U.S.), which used complete RPMI medium and added mouse SCF (30ng / mL, BioSource, Camarillo, CA ), mouse Flt3L (50ng / mL, PeproTech, Rocky Hill, NJ), mouse IL-7 (0.5ng / mL, PeproTech), indomethacin (2g / mL, Sigma), gentamicin (20g / mL) and 10% fetal bovine serum, the inoculation concentration is 2X10 6 cells / well. Incubate the cells at 37 °C and 5% CO 2 cultured in an incubator for 6 days. After 3 days, half of the culture supernatant was discarded and replaced with fresh new medium having the same composition as above. After further incubation for 6 days, CD122 was isolated using FITC-conjugated anti-CD122 and anti-FITC antibodies coupled to MACS magnetic beads + Immature NK cells (hereinafter referred to as "pNK cells"). The purity of the pNK cells was determined by FACS and a purity higher than...

Embodiment 3

[0071] Example 3: Expression of stage-specific VDUP1 gene during differentiation of isolated and purified NK cells

[0072] To obtain stage-specific NK developmental cells, Lin was isolated from mouse bone marrow - c-kit + HSCs (>95%) were then cultured for 6 days in the presence of SCF, Flt-3L and IL-7. CD122 was isolated + pNK cells (95%) followed by FACS analysis. pNK cells were cultured for 6 days in the presence of IL-15 alone (-OP9) or IL-15 and OP9 stromal cells together (+OP9), followed by FACS analysis. When the cells were co-cultured with OP9 stromal cells, the number of mNK cells increased even more (-OP9; 94% and +OP9; >95%). LY49 receptors on the surface of mNK cells play an important role in mNK cell function, and their expression is regulated by signal transduction that communicates with other immune cells. To investigate whether the co-culture of bone marrow-derived HSCs and stromal cells is necessary for the expression of the LY49 receptor, the NK recepto...

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 present invention is related to an agent for differentiating hematoietic stem cell into natural killer cell comprising VDUP1 protein or gene encoding the same, and a method of differentiating hematopoietic stem cell into natural killer cell using thereof. The present invention reveals for the first time that the VDUP1 gene is a critical factor for the regulation of differentiation of natural killer cell by generating a mouse deficient in VDUP1 gene, which confirms that VDUP1 gene is required for NK maturation. Thus, by the regulation of VDUP1 gene, the modulation of NK cells that have ability to kill cancer cells is possible and can be utilized for cell therapeutics.

Description

technical field [0001] The present invention relates to a reagent for differentiating hematopoietic stem cells into natural killer cells, and a method for using the reagent to differentiate hematopoietic stem cells into natural killer cells, more specifically, to a reagent for differentiating hematopoietic stem cells into natural killer cells, the reagent containing as VDUP1 (vitamin D3 upregulating protein 1) or gene encoding it or vitamin D3 regulating VDUP1 as an effective ingredient, and a method for differentiating HSCs into NK cells using the agent. Background technique [0002] Hematopoietic stem cells, a type of stem cells, are capable of differentiating into various blood components (red blood cells or erythrocytes, white blood cells or leukocytes, platelets, and lymphocytes) by any chance, and are constantly autologous in the body Regenerate and differentiate into immune cells. Among the cells forming the immune system, natural killer cells (hereinafter referred t...

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 Applications(China)
IPC IPC(8): C12N5/00C12N5/08C12N5/0783
CPCC07K14/4703C12N2501/998C12N5/0646
Inventor 崔仁杓尹锡兰李基宁金善昱柳大烈
Owner 医疗细胞公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com