Cell factor for amplifying umbilical cord blood hematopoietic stem cells and culture medium thereof

A technology of hematopoietic stem cells and cytokines, which is applied in the direction of cell culture active agents, cytokines/lymphokines/interferons, blood/immune system cells, etc., can solve the problem of low amplification factor of umbilical cord blood hematopoietic stem cells and failure to maintain stem cell characteristics and other issues to achieve the effect of increasing the amplification factor

Active Publication Date: 2014-01-15
广东昭泰细胞生物科技有限公司
View PDF4 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the defects in the prior art that the in vitro expansion of umbilical cord blood hematopoietic stem cells is not high and the characteristics of stem cells cannot be maintained, the present invention discloses a cytokine and its cul

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
  • Cell factor for amplifying umbilical cord blood hematopoietic stem cells and culture medium thereof
  • Cell factor for amplifying umbilical cord blood hematopoietic stem cells and culture medium thereof
  • Cell factor for amplifying umbilical cord blood hematopoietic stem cells and culture medium thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: A cytokine for expanding umbilical cord blood hematopoietic stem cells developed by the present invention, the cytokine is obtained by the following preparation method, which includes the following steps, (1) from a mouse fetus with a gestational age of 11 days Fetal liver stromal cells were isolated from the liver; fetal liver is the hematopoietic organ with the strongest vitality in mammalian individual development, and fetal liver contains stromal cells that support the proliferation of hematopoietic stem cells; fetal liver stromal cells will adhere to the wall when cultured in vitro;

[0023] (2) Fetal liver stromal cells isolated from step (1), transforming these primary stromal cells with SV40 large T antigen, and then continuously passaged for more than 30 times to obtain immortalized stromal cells;

[0024] (3) Pick 100 individual stromal cells from the immortalized stromal cells established in step (2) and expand and culture them respectively to obtai...

Embodiment 2

[0042] Example 2: A cytokine for expanding umbilical cord blood hematopoietic stem cells developed by the present invention, the cytokine is obtained by the following preparation method, which includes the following steps, (1) from a mouse fetus with a gestational age of 11 days Fetal liver stromal cells were isolated from the liver; fetal liver is the hematopoietic organ with the strongest vitality in mammalian individual development, and fetal liver contains stromal cells that support the proliferation of hematopoietic stem cells; fetal liver stromal cells will adhere to the wall when cultured in vitro;

[0043] (2) Fetal liver stromal cells isolated from step (1), transforming these primary stromal cells with SV40 large T antigen, and then continuously passaged for more than 30 times to obtain immortalized stromal cells;

[0044] (3) Pick 100 individual stromal cells from the immortalized stromal cells established in step (2) and expand and culture them respectively to obtai...

Embodiment 3

[0052] Example 3: A cytokine for expanding umbilical cord blood hematopoietic stem cells developed by the present invention, the cytokine is obtained by the following preparation method, which includes the following steps, (1) from a mouse fetus with a gestational age of 11 days Fetal liver stromal cells were isolated from the liver; fetal liver is the hematopoietic organ with the strongest vitality in mammalian individual development, and fetal liver contains stromal cells that support the proliferation of hematopoietic stem cells; fetal liver stromal cells will adhere to the wall when cultured in vitro;

[0053] (2) Fetal liver stromal cells isolated from step (1), transforming these primary stromal cells with SV40 large T antigen, and then continuously passaged for more than 30 times to obtain immortalized stromal cells;

[0054] (3) Pick 100 individual stromal cells from the immortalized stromal cells established in step (2) and expand and culture them respectively to obtai...

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 discloses a cell factor, the cell factor is prepared through the following steps: (1) processing primary stroma cells; (2) obtaining immortalized cells; (3) subjecting the fetal hepatocytes selected in the step (2) to continuous subculture to obtain a cell line; (4) obtaining a total RNA; (5) carrying out an inverse transcription process to convert the total RNA into cDNA; (6) carrying out a gene chip analysis; (7) selecting the high-expression gene sequence, which can promote the in-vitro amplification of umbilical cord blood hematopoietic stem cells, from the rat fetal hepatocytes; (8) retrieving the nucleotide sequence obtained in the step (7); (9) obtaining a 293T cell; (10) obtaining a corresponding protein of the 293T cell; (11) adding the purified protein obtained in the step (10) into a StemSpan culture medium containing SCF, TPO, FLT3L, and PTN, and testifying the promotion effect on the in-vitro amplification of umbilical cord blood hematopoietic stem cells. The cell factor has the advantages of prominent increasing on amplification multiple of umbilical cord blood hematopoietic stem cells and maintenance of stem cell characteristics.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a protein for expanding umbilical cord blood hematopoietic stem cells and a culture medium thereof. Background technique [0002] Hematopoietic stem cells are widely used in the treatment of leukemia, malignant tumors, severe non-malignant hematological diseases and some genetic diseases, because these cells have the ability of self-renewal, high reproduction, and differentiation in multiple directions, which also means Hematopoietic stem cells have great application value in the clinical treatment of malignant blood diseases. [0003] Currently, there are three main sources of hematopoietic stem cells for transplantation: bone marrow, peripheral blood and umbilical cord blood. Bone marrow and peripheral blood are currently widely used sources of hematopoietic stem cell transplantation, but there are problems such as difficulty in matching, and the collection process will bring pain...

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
IPC IPC(8): C07K14/52C12N15/19C12N15/85C12N5/0789
CPCC07K14/52C12N5/0647C12N2500/30C12N2501/20
Inventor 李鹏肖毅仁蒋志武赖允鑫姚瑶
Owner 广东昭泰细胞生物科技有限公司
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