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Tissue engineering lymph node model and construction method thereof

A technology of tissue engineering and construction method, applied in medical science, prosthesis, etc., to achieve the effect of simple operation process and mild implementation conditions

Active Publication Date: 2011-11-02
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there is no report on a tissue engineered lymph node model based on collagen-Mstrigel scaffold material

Method used

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  • Tissue engineering lymph node model and construction method thereof
  • Tissue engineering lymph node model and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Using human peripheral blood mononuclear cells as seed cells to construct a tissue engineered lymph node model

[0028] (1) Separation of human peripheral blood mononuclear cells by density gradient centrifugation

[0029] The heparinized anticoagulated blood was diluted one-fold with Hanks solution, spread lightly on Ficoll separation medium (specific gravity 1.077g / mL), and then centrifuged horizontally at 2000rpm for 30min. Mononuclear cells are suspended in the upper layer of the separation medium and appear as a white film. Carefully draw the mononuclear cell layer with a pipette, add Hanks solution and wash 3 times. Add RPMI-1640 medium containing 10% fetal bovine serum for routine culture.

[0030] (2) Isolation and culture of primary mouse fibroblasts (MEFs)

[0031] The embryonic tissues of dead pregnant mice were taken, transferred to culture flasks, and washed with PBS. Pour off the PBS, add an appropriate amount of trypsin solution for digestio...

Embodiment 2

[0040] Example 2 Using mouse bone marrow mononuclear cells as seed cells to construct a tissue engineered lymph node model

[0041] (1) Separation of mouse bone marrow mononuclear cells by density gradient centrifugation

[0042] Take the femur of the dead mouse, put the femur into sterile cold PBS; use a 1mL syringe to flush out the cells in the mouse femur until the femoral cavity is white; mix the cells well, use a 200-mesh filter to filter, collect and filter Liquid in a 10mL centrifuge tube, 1500rpm. Centrifuge for 5 minutes; discard the supernatant, add 8 mL of NH4Cl-Tris solution, suspend the cells, dissolve the red blood cells for about 6 minutes, centrifuge at 1500 rpm for 5 minutes. Discard the supernatant, add 5mL sterile cold PBS to the centrifuge tube, and resuspend the cells; gently add the above cell suspension to 5mL Ficoll-paqul (1.086g / mL), centrifuge at 1500rpm for 30min; carefully take out the centrifuge tube, the solution Divided into three layers: trans...

Embodiment 3

[0053] Example 3 Using mouse spleen lymphocytes as seed cells to construct a tissue engineered lymph node model

[0054] (1) Separation of mouse spleen lymphocytes by density gradient centrifugation

[0055] The spleen tissue of the dead mouse was taken, placed in sterile cold PBS, then the spleen was cut into pieces, filtered through a 200-mesh filter, and the filtrate was collected in a 10mL centrifuge tube at 1500rpm. Centrifuge for 5 minutes; discard the supernatant, add 8 mL of NH4Cl-Tris solution, suspend the cells, dissolve the red blood cells for about 8 minutes, centrifuge at 1500 rpm for 5 minutes. Discard the supernatant, add 5mL sterile cold PBS to the centrifuge tube, and resuspend the cells; gently add the above cell suspension to 5mL Ficoll (1.086g / mL), centrifuge at 2000rpm for 15min; carefully take out the centrifuge tube, and divide the solution into Three layers: from top to bottom are transparent PBS layer, mononuclear cell layer, and RBS layer; use a thin...

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Abstract

The invention discloses a tissue engineering lymph node model and a construction method thereof and belongs to the technical field of tissue engineering and regenerative medicine. The tissue engineering lymph node model comprises lymphocytes separated from lymphoid tissues and a collagen-Matrigel scaffold. The construction method for the tissue engineering lymph node model comprises the following steps of: preparing lymphocyte suspension and primary generation mouse fibroblast cell suspension; and compounding mixed cell suspension and the collagen-Matrigel scaffold, and culturing in a mold subjected to static stretching to obtain the tissue engineering lymph node model. The scaffold system subjected to static stretching can better simulate in-vivo environment and support the growth and proliferation of the lymphocytes, secretion of cell factors and the expression of surface markers, and has important significance for applying the tissue engineering lymph node model to immunotoxicity evaluation of biological materials, immune medicine screening and the like. The construction method has the advantages of simple operating process and mild implementation conditions.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering and regenerative medicine, and in particular relates to a tissue engineering lymph node model and a construction method thereof. Background technique [0002] The importance of the lymphatic system to the human body is becoming increasingly apparent, and the need for the development of lymphoid tissue biology research promotes the research on the construction of suitable models. Tissue-engineered lymphoid organs (lymph nodes, spleen, lymph vessels, and lymphocytes) can not only be used as in vitro research models, but also have potential application value for transplantation to treat lymphoid defects caused by diseases or injuries. Chistoph Giese et al. (Chistoph Giese et al., Artificial Organs, 2006, 30, 803) succeeded in cultivating dendritic cells derived from peripheral blood mononuclear cells in a bioreactor in vitro, and Sachiko Suematsu et al. (Sachiko Suematsu et al., Nuture Bio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/24
Inventor 王常勇郝彤周瑾王妍林秋霞杜芝燕段翠密
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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