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Construction method of stem cell bank for preventing and treating novel coronavirus pneumonia

A technology of stem cell bank and construction method, which is applied in the field of construction of stem cell bank for the prevention and treatment of new coronary pneumonia, and can solve problems such as stem cell bank of new coronavirus infectious disease that has not been seen

Inactive Publication Date: 2021-04-09
翁炳焕
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no literature reports on the stem cell bank dedicated to the prevention and treatment of the new coronavirus infectious disease and its construction method

Method used

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  • Construction method of stem cell bank for preventing and treating novel coronavirus pneumonia
  • Construction method of stem cell bank for preventing and treating novel coronavirus pneumonia
  • Construction method of stem cell bank for preventing and treating novel coronavirus pneumonia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1. Preparation of recombinant DNA stem cells

[0040] 1. Collection of mesenchymal stem cells (MSCs)

[0041] 1.1. Collection of amniotic fluid mesenchymal stem cells

[0042] According to the prenatal diagnosis process, the amniotic fluid cells of the pregnant women to be tested are collected, cell culture and prenatal diagnosis are carried out, and the adherent growth spindle amniocytes or mesenchymal stem cells are screened from the remaining amniotic fluid cells under an inverted microscope.

[0043] 1.2. Collection of umbilical cord mesenchymal stem cells

[0044] Take out the frozen umbilical cord from the liquid nitrogen tank, thaw it quickly in a 37°C water bath, wash it with sterile PBS, remove the residual blood, and cut the umbilical cord into a size of about 1mm 3 The tissue block was placed in a petri dish covered with fetal bovine serum, and after 6 hours at 37°C, DMEM medium with 10% fetal bovine serum was added, at 37°C in 5% CO 2 Cultivate in...

Embodiment 2

[0188] Example 2. Application of DNA Recombinant Stem Cells in the Preparation of COVID-19 Vaccines

[0189] 1. RBD gene assembly of recombinant DNA stem cells

[0190] 1.1. Synthesis of RBD gene

[0191] The amino acid sequence of the S protein of SARS-CoV-2 was analyzed with reference to the GenBank accession number (MN908947.3). The RBD amino acids were Gly319~Asn541, and amino acid codon synonymous substitutions were used to synthesize the whole genome of the nucleic acid corresponding to the RBD peptide. . Codon optimization before synthesis, the purpose is to increase the G+C% content of the genome, increase the stability of mRNA in mammalian cells or the rate of mRNA input into the cytoplasm, avoid the loss of rare tRNAs, enhance protein expression, and improve immunogenicity . The specific method is as follows: Statistical analysis of the usage frequency of NC_045512.2 strain virus codons and human codons (http: www.kazusa.or.jp / codon) , using human preferred codo...

Embodiment 3

[0229] Example 3, Construction of recombinant DNA stem cell bank

[0230] 1. Construct DNA recombinant stem cells according to the present invention.

[0231] 2. Construct a DNA recombinant stem cell bank dedicated to the individualized treatment of COVID-19, such as Figure 5 According to the donor's name and / or tissue type, the recombinant DNA stem cells were stored in liquid nitrogen at -196°C.

[0232] 3. When an individual is infected with the new coronavirus, the stem cells of their own or the same type can be quantitatively taken out from the stem cell bank for in vitro expansion and culture for individualized treatment. During the outbreak of infectious diseases, stem cells of various types can be taken out for industrial expansion and culture at the same time, so that different individuals can choose stem cells of the same type for individualized treatment of COVID-19.

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Abstract

A construction method of a stem cell bank for preventing and treating novel coronavirus pneumonia comprises the following steps of: collecting residual amniotic fluid cells, newborn umbilical cord blood, umbilical cord and placenta tissues after prenatal diagnosis, carrying out stem cell induction, immortalized gene transfection, novel coronavirus targeted interference gene assembly, novel coronavirus susceptibility gene knockout or knock-in and novel coronavirus antibody generation gene assembly to prepare immortalized stem cells, DNA recombinant stem cells, RNA interference stem cells, gene silencing stem cells and various stem cell vector vaccines for preventing and treating novel coronavirus pneumonia, collecting the prepared stem cell products in -196 DEG C liquid nitrogen in a centralized mode according to donor names, ABO blood types or HLA types, and establishing a renewable special stem cell bank. When the novel coronavirus is epidemic, various types of stem cells or vaccines thereof can be extracted from the stem cell bank to be cultured and amplified, and individualized treatment is performed by the patient's own or homologous stem cells, so that side effects such as immunological rejection and the like of traditional stem cell or vaccine prevention and treatment are reduced, and the problem of lack of COVID-19 special-effect medicines and stem cells is solved.

Description

technical field [0001] The invention relates to a method for constructing a stem cell bank for the prevention and treatment of COVID-19, which is an infectious disease prevention and control technology in the field of biomedicine. Background technique [0002] The new coronavirus (SARS-CoV-2) has posed a serious threat to global public health. Due to the lack of effective antiviral drugs, the treatment of the new coronavirus disease (COVID-19) is still full of challenges, and it is urgent to explore new effective treatments , in order to improve the treatment rate of clinically critical patients. [0003] Mesenchymal stem cells (MSCs) are a group of pluripotent stem cells originating from the mesoderm, which have the potential of self-renewal, multilineage differentiation and immune regulation. As seed cells, MCS can be used to repair / replace various injured or diseased tissues and organs. A number of clinical studies on MSC have been carried out around the world to treat s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C40B50/06C40B40/02C12N15/867A61K35/28A61P31/14
CPCA61K35/28A61P31/14C07K14/005C12N9/1276C12N9/485C12N15/1131C12N15/86C12N2310/14C12N2710/22022C12N2740/15043C12N2800/107C12Y207/07049C12Y304/17023C40B40/02C40B50/06
Inventor 翁炳焕
Owner 翁炳焕
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