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Application of SUSD2 protein as marker

A protein and cell technology, applied in the field of SUSD2 protein as a marker, can solve problems such as limiting diabetes cell replacement therapy

Active Publication Date: 2014-12-17
PEKING UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, how to induce the differentiation of pancreatic precursor cells into functionally mature pancreatic beta cells in vitro greatly limits the realization of diabetes cell replacement therapy. In the process of pancreatic precursor cells differentiation into pancreatic beta cells, the correct pancreatic endocrine The realization of efficient induction of cell differentiation is the most important step

Method used

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  • Application of SUSD2 protein as marker
  • Application of SUSD2 protein as marker
  • Application of SUSD2 protein as marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1. Discovery and application of SUSD2 as marker gene of pancreatic endocrine precursor cells

[0046] 1. SUSD2 as a marker gene of pancreatic endocrine precursor cells differentiated from modified human pluripotent stem cells

[0047] 1.1. Obtaining pancreatic endoderm cells

[0048] The EGFP gene (NC_013179.1(3313..4126)) is located in Chromosome 10 through homologous recombination (Ngn3 contig number: NT_030059.13, the position of the left homology arm in the genome: 71325654-71332154; the right homology The position of the arm in the genome: 71332158-71467568) the isolated human pluripotent stem cell line H1 (WiCell Research Institute, NIH number is WA01), and the modified human pluripotent stem cell line NGN3-EGFP was obtained. Inoculate it into cell culture medium I at 15%-20%, and cultivate for 1 day; then directly replace with cell culture medium II, and cultivate for 1-2 days; then directly replace with cell culture medium III, and cultivate for 1 day; ...

Embodiment 2

[0126] Example 2, Expression of SUSD2 Gene Sorting Human Pancreatic Endocrine Precursor Cells or Their Progeny Cells That Don’t Secrete Hormone

[0127] 1. Obtaining pancreatic endoderm cells

[0128] Differentiate the isolated human pluripotent stem cell line according to the method in 1.1 to obtain pancreatic endoderm cells;

[0129] 2. Detection of SUSD2 gene expression by flow cytometry and sorting of human pancreatic endocrine precursor cells or their progeny cells that do not secrete hormones

[0130] Pancreatic endoderm cells were detected by flow cytometry, and the antibody used was SUSD2 direct-labeled antibody (PE-labeled anti-SUSD2 monoclonal antibody derived from mouse was purchased from BioLegend, the product catalog number was 327406), stained, and then stained with fluorescent II Anti counterstain.

[0131] If some cells express the SUSD2 gene, some cells are or are candidates for human pancreatic endocrine precursor cells or their non-hormone-secreting progen...

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Abstract

The invention discloses an application of SUSD2 protein as a marker and particularly provides an application of the SUSD2 protein as the marker in identification, selection or separation of pancreas internal secretion precursor cells and / or newborn pancreas internal secretion cells, and an application of an mRNA, for encoding the SUSD2 protein, of a precursor protein as the marker in identification of the pancreas internal secretion precursor cells and / or the newborn pancreas internal secretion cells. In the invention, through analysis of gene expression of pancreas entoderm cells sourced by induced directional differentiation of human pluripotent stem cells, enrichment expression of a SUSD2 gene in the ancreas internal secretion precursor cells and the newborn pancreas internal secretion cells is found. In addition, a protein encoded by the SUSD2 gene is a receptor protein on cell membranes. With the protein as the marker, the identification, the selection or the separation of the pancreas internal secretion precursor cells and the newborn pancreas internal secretion cells can be carried out, which has an important significance on research of pancreas relative cells in various growing periods.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of SUSD2 protein as a marker. Background technique [0002] Pancreatic beta cells derived from human pluripotent stem cells can provide a sufficient source of donor islet cells for cell replacement therapy of diabetes, especially type I diabetes. In addition, the directed differentiation of human pluripotent stem cells into pancreatic beta cells can also provide an in vitro research model for studying the development of human pancreas. By simulating the development process in vivo, the directed differentiation of human pluripotent stem cells to pancreatic beta cells mainly includes the following steps: intraembryonic endoderm, gut tube, posterior foregut, pancreatic endoderm, and pancreatic endocrine cells. Among them, the cells in the pancreatic endoderm stage are mixed cells, usually including pancreatic precursor cells, pancreatic endocrine precursor cells, and new pancre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/6883G01N33/68C12Q2600/158C12Q2600/156G01N33/507G01N33/5073G01N2500/10G01N33/56966C12Q1/6886G01N33/53G01N33/54326G01N33/577G01N33/6872G01N33/582G01N2333/705
Inventor 邓宏魁刘海松朱迪聪杨欢梁振
Owner PEKING UNIV
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