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Construction method and application of research model of bystander effector damaged hematopoietic stem progenitor cells

A bystander effect and research model technology, applied in the field of research model construction, can solve the problems of long-term hematopoietic reconstruction ability, self-renewal ability and decline of hematopoietic progenitor cell clone formation ability in vitro

Pending Publication Date: 2022-01-07
HAIHE LAB OF CELL ECOSYSTEM +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Decreased long-term hematopoietic reconstitution, self-renewal, and in vitro clonogenicity of hematopoietic progenitor cells (HPCs) in HSCs

Method used

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  • Construction method and application of research model of bystander effector damaged hematopoietic stem progenitor cells
  • Construction method and application of research model of bystander effector damaged hematopoietic stem progenitor cells
  • Construction method and application of research model of bystander effector damaged hematopoietic stem progenitor cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0136] In this example, an in vivo research model of bystander effect damage to hematopoietic stem and progenitor cells and an in vivo research model of bystander effect damage to hematopoietic stem and progenitor cells intervened by antioxidant drugs were constructed. The steps are as follows:

[0137] (1) Female immunodeficiency mice (NOG mice) aged 6 to 8 weeks and weighing 18 to 22 g were randomly divided into NR group (control group), IR group (X-ray irradiation group), IR+NAC group group (pretreatment with NAC before X-ray irradiation), IR+SF group (pretreatment with Sulforaphane before X-ray irradiation), and IR+Res group (pretreatment with Resveratrol before X-ray irradiation);

[0138] (2) Mice in IR+NAC group, IR+SF group and IR+Res group were treated with antioxidant drugs 7 days before irradiation, and the treatment plan was as follows:

[0139] IR+NAC group: intraperitoneal injection, 100mg / kg / day, once a day, for 7 consecutive days, and at the same time fed water...

Embodiment 2

[0157] In this example, an in vitro research model of bystander effect damage to hematopoietic stem and progenitor cells and an in vitro research model of bystander effect damage to hematopoietic stem and progenitor cells intervened by antioxidant drugs were constructed. The steps are as follows:

[0158] (1) Configure complete medium for stem cells (fetal bovine serum containing 10% volume fraction, 50ng / mL TPO, 50ng / mL SCF and 50ng / mL Flt-3, the balance being IMDM medium), and containing NAC (100μM), Stem cell complete medium of SF (2.5 μM) and Res (0.1 μM);

[0159] (2) Isolate human bone marrow cells, lyse the red blood cells, and divide them into NR group (control group), IR group (X-ray irradiation group), IR+NAC group (NAC pretreatment before X-ray irradiation), IR+SF group (pretreatment with Sulforaphane before X-ray irradiation) and IR+Res group (pretreatment with Resveratrol before X-ray irradiation), 30 minutes before irradiation, bone marrow cells were resuspended ...

Embodiment 3

[0165] In this embodiment, the human CD34 of the IR group and the NR group in the embodiment 1 subjected to RIBE + The cells were transplanted into mice, and the implantation and differentiation of the cells were detected. The steps are as follows:

[0166] (1) Female NOG mice aged 6 to 8 weeks and weighing 18 to 22 g were randomly divided into groups, and the mice were irradiated with 2Gy of X-rays at a dose rate of 1.2Gy / min the day before transplantation;

[0167] (2) The human CD34 of each group obtained by sorting + Cells were transplanted into mice irradiated with 2Gy through the tail vein, and human CD45 in peripheral blood of mice was continuously detected 4 to 20 weeks after transplantation. + cell ratio;

[0168] (3) 20 weeks after transplantation, the mice were killed by decapitation, and the bone marrow cells of the mice were collected 1×10 6 After washing with staining buffer, resuspend, add antibody, incubate at 4°C in the dark for 30 min, wash once with 2 mL ...

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Abstract

The invention provides a construction method and application of a research model of bystander effector damaged hematopoietic stem progenitor cells. The construction method comprises a construction method of an in-vivo research model of the bystander effector damaged hematopoietic stem progenitor cells, the constructed in-vivo research model of the bystander effector damaged hematopoietic stem progenitor cells, an in-vitro research model of the bystander effector damaged hematopoietic stem progenitor cell and the constructed in-vitro research model of the bystander effector damaged hematopoietic stem progenitor cell. The invention further provides application of the N-acetylcysteine, the sulforaphane and the resveratrol in relieving and / or treating bystander effect damage of the hematopoietic stem progenitor cells and improving the hematopoietic stem progenitor cell transplantation effect. The application proves that the generation of active oxygen is the main mechanism of HSPC damage caused by RIBE, and the antioxidant drug can reduce the damage of RIBE to HSPC, thereby providing a theoretical basis for the formulation of a new strategy for HSC transplantation.

Description

technical field [0001] The invention belongs to the technical field of hematopoietic stem cell transplantation, and in particular relates to a construction method and application of a research model of hematopoietic stem and progenitor cells damaged by the bystander effect. Background technique [0002] Hematopoietic stem cell (HSC) transplantation is an important means to treat various malignant hematological diseases and immune dysfunction. The low rate of HSC transplantation and slow hematopoietic reconstitution are the bottlenecks in the application of HSC in the treatment of clinical diseases. At present, there is no effective strategy to significantly increase the rate of HSC transplantation and the speed of hematopoietic reconstitution. The fundamental reason is that the molecular mechanism of HSCs affected by the recipient microenvironment and HSC regeneration after transplantation is still unclear. [0003] At present, when HSC transplantation is performed clinical...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789C12N5/077A01K67/027A61K31/26A61P7/00A61K31/05A61K31/198
CPCC12N5/0647A01K67/027A61K31/198A61K31/26A61K31/05A61P7/00C12N2502/1394C12N2529/10C12N2501/145C12N2501/125C12N2501/26A01K2207/35A01K2207/12A01K2227/105A01K2267/0331A61K2300/00
Inventor 胡林萍程涛尹秀秀姜尔烈庞爱明张雅文
Owner HAIHE LAB OF CELL ECOSYSTEM
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