Adult sertoli cell-pancreas islet compound and preparation method for same

A technology of testicular Sertoli cells and complexes, applied in artificial cell constructs, animal cells, vertebrate cells, etc., can solve the problems of immediate restoration of blood supply, destruction of microcirculation system, nutritional deficiency damage, etc., to improve islet nutritional deficiencies Status, prolong survival time, suppress rejection effect

Inactive Publication Date: 2013-04-24
THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
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Problems solved by technology

[0003] The islet isolation and purification process destroys its internal microcirculatory system, resulting in that the blood supply cannot be restored immedia

Method used

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  • Adult sertoli cell-pancreas islet compound and preparation method for same
  • Adult sertoli cell-pancreas islet compound and preparation method for same
  • Adult sertoli cell-pancreas islet compound and preparation method for same

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Embodiment Construction

[0024] The present invention will be described in more detail below.

[0025] Step 1: Adult Sertoli cells were isolated, purified and cultured. Wash the testis with Hank's solution at 4°C, remove the capsule and blood vessels, then pick out the seminiferous tubules with ophthalmic forceps, try to peel off the interstitial tissue around the seminiferous tubules and cut them into pieces, put the testicular tissue in 2.5 g / L trypsin, Digest with 0.4 g / L DNase I in a constant temperature water bath at 37°C for 30 min, stop digestion in DMEM / F12 medium containing 100 mL / L fetal bovine serum, centrifuge to remove supernatant, 5 g / L collagenase V, 1 g / L Digest L hyaluronidase in a constant temperature water bath at 37°C for 20 min, stop digestion with DMEM / F12 medium containing 100 mL / L fetal bovine serum, filter through a 100-mesh sieve, and wash 3 times with serum-free DMEM / F12 medium Afterwards, centrifuge to remove the supernatant. The obtained pellets were resuspended in DMEM / ...

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Abstract

The invention discloses an adult sertoli cell-pancreas islet compound and a preparation method for the same, which solve the problem of influence on pancreas islet transplantation effect due to the transplant dysfunction because the pancreas islet separated and purified presently generates hypoxic-ischemic and immunological rejection reactions after being transplanted and then loses the function. The preparation method comprises the following steps of: 1, separating and purifying adult pancreas islet by adopting a semi-automatic filling and continuous density gradient methods; 2, separating and purifying adult sertoli cells by adopting a two-step digestion method; and 3, constructing the pancreas islet-sertoli cell compound by adopting a specific co-culture method. The adult sertoli cell-pancreas islet compound and the preparation method for the same disclosed by the invention have the advantage that pancreas islet cells and sertoli cells are coupled into a compound, the compound is used as a transplant, and the sertoli cells are attached to the surface of the pancreas islet, so as to improve the nutritional status of the pancreas islet, inhibit rejection reaction, and increase the survival rate of the pancreas islet; the operation is simple and convenient; and the success rate is high.

Description

technical field [0001] The invention designs a preparation method of an adult pancreas islet transplantation complex. Background technique [0002] Islet transplantation has become the most promising cure for type 1 diabetes, but donor shortages and graft loss limit its widespread use. [0003] The islet isolation and purification process destroys its internal microcirculatory system, resulting in that the blood supply cannot be restored immediately after transplantation like pancreas transplantation, but it takes a long time to rebuild the blood supply system, resulting in nutritional deficiency damage. In addition, immune rejection is still an important problem that plagues the widespread clinical development of islet transplantation. It is an ideal method to induce immune tolerance of islet transplantation with cells with immune regulation function. Contents of the invention [0004] Sertoli cells have both nutritional support and immune regulation functions. Based on...

Claims

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Application Information

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IPC IPC(8): A61K35/48A61K35/39C12N5/071A61P5/48A61P37/08
Inventor 李杨薛武军丁小明冯新顺田小辉田普训丁晨光
Owner THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
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