Preparation method of enhanced green fluorescence SCID mice model and model application

A green fluorescence and mouse model technology, which is applied in animal husbandry and other fields, can solve the problems that human tissue cells cannot be studied with fluorescent tracer experiments, and achieve high xenotransplantation efficiency

Inactive Publication Date: 2017-11-21
THE SECOND HOSPITAL AFFILIATED TO SUZHOU UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Because the tissues and organs of the whole body can emit green fluorescence, it is suitable for the in vivo study of the interaction between exogenous tissues or cells (usually another fluorescent marker) and host (mouse) tissue cells in the mouse body, but the small Due to the sound immune function of mice, human tissues or cells cannot be transplanted and survived in their bodies, so they cannot be used for in vivo fluorescent tracer experiments on human tissue cells

Method used

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  • Preparation method of enhanced green fluorescence SCID mice model and model application
  • Preparation method of enhanced green fluorescence SCID mice model and model application
  • Preparation method of enhanced green fluorescence SCID mice model and model application

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Example 1 Preparation of NOD / SCID / EGFP mice (enhanced green fluorescent SCID mouse model)

[0021] 1. Main materials and equipment:

[0022] NOD / SCID and green fluorescent protein transgenic mice C57BL / 6-Tg (CAG-EGFP) were purchased from the Model Animal Research Institute of Nanjing University (SCXK Su 2010-0001); independent ventilation cages were purchased from Suzhou Fengshi Experimental Animal Equipment Co., Ltd. ( IVC-II); the biological safety cabinet was purchased from Singapore Yisi Cleaning Equipment Co., Ltd. (ESCO); the small animal multi-mode live imager (In-vivo imaging system FX Pro) was purchased from Kodak Company. Fluorescence flashlight was purchased from Nightsea Company, PCR instrument (AB Company), flow cytometer (BD Company), fluorescence inverted microscope (Nikon Company), CO2 incubator (Germany Heraeus Company). SLY series animal brain slice molds were purchased from Beijing Shuolinyuan Technology Co., Ltd. The frozen microtome was purchased ...

Embodiment 2

[0030] Example 2 Application of Enhanced Green Fluorescent SCID Mouse Model

[0031] The enhanced green fluorescent SCID mouse model obtained by using the method in Example 1 can be used for the research of breast cancer brain metastasis model.

[0032] 1. Clinical specimen tissue blocks were directly inoculated into NOD / SCID / EGFP mice

[0033] Clinical specimen information: Human breast cancer slices were obtained from a 40-year-old married female patient undergoing surgical treatment, provided by the Pathology Department of the Second Affiliated Hospital of Soochow University. Later, due to brain metastases, he was admitted to neurosurgery to remove brain metastases.

[0034] Tissue inoculation method: Select 5 SCID female mice aged 4-6 weeks and expressing EGFP, disinfect the skin of the head after anesthesia with chloral hydrate, cut the skin longitudinally at the center of the top of the head 0.5cm, expose the skull, place it 1.0mm in front of the bregma, At 2.5mm to th...

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Abstract

The invention discloses a preparation method of an enhanced green fluorescence SCID mice model. The method comprises the following steps: adopting the mode of crossing and backcrossing among multiple generations of C57BL / 6-Tg (CAG-EGFP) mice and NOD / SCID mice and screening the multiple generations to establish steadily hereditary NOD / SCID / EGFP mice systemically expressing green fluorescence, namely, an enhanced green fluorescence SCID mice model sheet. The preparation method employs the breeding method featuring crossing and backcrossing among multiple generations to raise NOD / SCID mice expressing enhanced green-florescent protein genes. Compared with nude mice, the SCID mice have more severe immune deficiency including combined T and B cell deficiencies. The invention further discloses an application of the enhanced green fluorescence SCID mice model. In terms of xenotransplantation study, the SCID mice are more efficient in xenotransplantation than the nude mice with deficient t lymphocytes. The method of the model is an excellent model for fluorescent animals with immune deficiencies.

Description

technical field [0001] The invention belongs to the field of animal models used in the medical field, in particular to mouse models. Background technique [0002] Fluorescent SCID mice, SCID (Severe Combined Immnue Deficiency) mice are severe combined immunodeficiency mice, successfully bred by American scholar Bosma[10] in 1983, it is a kind of innate T, B lymphocyte double immune Deficient animals can thus be good recipients of xenografts. The emergence of NOD-SCID mice in 1992 further reduced the immunity of mice and improved the transplantation efficiency of human peripheral blood mononuclear cells and hematopoietic stem cells. [0003] Over the past three decades, advancements in cell engraftment and related studies of human peripheral blood mononuclear cells (PBMCs) and hematopoietic stem cells (HSCs) in severe combined immunodeficiency (SCID) mice have driven advances in hematology, infection Developments in related fields such as disease, autoimmunity, and tumor im...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/02
CPCA01K67/02
Inventor 董军陈延明盛敏峰薛彦平刘加驰黄强
Owner THE SECOND HOSPITAL AFFILIATED TO SUZHOU UNIV
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