Stem cell biological product for treating liver cirrhosis and preparation method and application thereof

A biological product and stem cell technology, applied in the field of stem cells, can solve problems such as insufficient effect, and achieve the effects of increasing the number, improving the therapeutic effect, and improving the specific homing ability

Pending Publication Date: 2020-04-03
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD +1
View PDF3 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the problem of insufficient effect of existing conventional treatment methods for liver cirrhosis, the prese...

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
  • Stem cell biological product for treating liver cirrhosis and preparation method and application thereof
  • Stem cell biological product for treating liver cirrhosis and preparation method and application thereof
  • Stem cell biological product for treating liver cirrhosis and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0038] Another embodiment of the present invention provides a method for preparing stem cell biological products as described above, comprising the following operations:

[0039] Obtain the CCR9 target gene;

[0040] constructing a vector comprising the CCR9 gene sequence;

[0041] obtain mesenchymal stem cells;

[0042] Vector transfected mesenchymal stem cells.

[0043] In some embodiments, the "obtaining the CCR9 gene of interest" includes the following operations:

[0044]Liver tissue samples were collected from patients, total RNA was extracted, amplification primers were designed, and the full-length CCR9 gene was amplified by reverse transcription-polymerase chain reaction method, followed by electrophoresis, and the target gene fragment of CCR9 was purified.

[0045] In some embodiments, the enzyme cleavage sites of the CCR9 gene sequence are Kpn1 and EcoR1, and Primer6.0 software is used to design the amplification primers of the CCR9 gene, and the primer sequences...

Embodiment 1

[0072] This example is used to illustrate the stem cell biological product disclosed in the present invention and its preparation method, including the following steps:

[0073] Step 1. Construction of the recombinant plasmid vector carrying the target gene CCR9

[0074] Step 1, obtaining the target gene. Collect liver tissue samples from patients, use Trizol reagent to extract total RNA, and identify whether total RNA is degraded by agarose electrophoresis. Design the amplification primers as:

[0075] CCR9-Kpn1-F1: ggggtaccat gacacccaca gacttcaca (SEQ ID NO.1)

[0076] CCR9-EcoR1-R1: cggaattctc agagggagag tgctcctg (SEQ ID NO. 2)

[0077] 1 μg of total RNA was used to reverse-transcribe cDNA with OligodT(20), and the full-length CCR9 gene was amplified by RT-PCR.

[0078] PCR conditions: Denaturation at 98°C for 15s, annealing and extension at 68°C for 3min, cycle 35 times, extension at 72°C for 15min after the last cycle. Agarose gel electrophoresis observation, gel cut...

Embodiment 2

[0094] This example is used to illustrate the stem cell biological product disclosed in the present invention and its preparation method, including most of the operating steps in Example 1, the difference being:

[0095] In step one, the amplification primers designed in step 1 are:

[0096] CCR9-Kpn1-F2:ggggtaccaa tgacacccac agacttcaca (SEQ ID NO.3)

[0097] CCR9-EcoR1-R1: cggaattctc agagggagag tgctcctg (SEQ ID NO. 4);

[0098] In step 3, the target fragment CCR9 is linked with the commercial plasmid vector pcDNA3.1-HisA, B, and C.

[0099] Construct pcDNA3.1-HisA, B, C-CCR9 expression vector.

[0100] In Step 3, umbilical cord mesenchymal stem cells are transfected with pcDNA3.1-HisA, B, C-CCR9.

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

In order to overcome a problem that an existing conventional treatment method for liver cirrhosis has a poor effect, the invention provides a stem cell biological product. The stem cell biological product comprises mesenchymal stem cells, wherein the mesenchymal stem cells are transfected into a vector, and the vector comprises a CCR9 gene sequence. Furthermore, the invention also discloses a preparation method and application of the stem cell biological product. The stem cell biological products utilizes that combination of CCL25 and receptor CCR9 has an effect of cell chemotaxis, improves aspecific homing ability of the mesenchymal stem cells at a focal part of liver cirrhosis, increases number of local stem cells, and effectively improves a therapeutic effect of the mesenchymal stem cells on liver cirrhosis.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and in particular relates to a stem cell biological product for treating liver cirrhosis and its preparation method and application. Background technique [0002] Liver cirrhosis is a common clinical chronic progressive liver disease. It is a chronic liver disease characterized by diffuse liver fibrosis, pseudolobule and regenerative nodule formation caused by one or more causes of long-term or repeated action. In my country, most of them are post-hepatitis cirrhosis, and a small part are alcoholic cirrhosis and schistosome cirrhosis. The peak age of onset of this disease is 35-48 years old, and the ratio of male to female is about (3.6-8):1. [0003] Studies have shown that the chemokine CCL25 is expressed in normal liver tissue, liver fibrosis, and liver tissue of patients with liver cirrhosis. As the course of liver cirrhosis progresses, the content of CCL25 in the liver tissue of patient...

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): A61K35/28A61K48/00A61P1/16C12N15/85
CPCA61K35/28A61K48/0041A61P1/16C12N15/85C12N2800/107
Inventor 曾宪卓余振东汪月王琳
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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