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Separating and purifying liquid for separation and purification of pancreatic islet during pancreatic islet transplantation and preparation method thereof

A technology for separation, purification and islet transplantation, which is applied in the fields of biology and medicine, can solve the problems of low purification efficiency and poor protection of islets, and achieve the effects of strong immunogenicity, reducing free radical damage, and reducing shear force

Inactive Publication Date: 2014-08-06
SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of islet isolation and purification, there is still no special separation medium for islet isolation and purification. The use of UW liquid and other organ preservation solutions still faces problems such as low purification efficiency and poor protection of islets. Therefore, it is urgent to develop a Islet separation and purification solution with high separation efficiency, strong islet protection and low cost

Method used

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  • Separating and purifying liquid for separation and purification of pancreatic islet during pancreatic islet transplantation and preparation method thereof
  • Separating and purifying liquid for separation and purification of pancreatic islet during pancreatic islet transplantation and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0027] The composition of each 1000ml islet isolation and purification solution is shown in Table 1, and the balance is water for injection.

[0028] Table 1

[0029] lactobionic acid 35.83g L-histidine 4.66g Raffinose 17.84g NaOH 4.60g KOH 0.39g CaCl 2 2H 2 o 0.0735g MgSO 4 ·7H 2 o 1.23g NaH 2 PO 4 3.45g polyethylene glycol-20 30.00g Glutathione 0.92g Allopurinol 0.14g Adenosine 1.34g Ulinastatin 100,000 units

[0030] Specific preparation method:

[0031] First, 0.294g of CaCl 2 2H 2 O dissolved in 20ml of water for injection; 1.23g of MgSO 4 ·7H 2 O was dissolved in 10ml of water for injection; 4.66g of L-histidine was dissolved in 200ml of water for injection; 30g of polyethylene glycol-20 was heated and dissolved in 200ml of water for injection, ready for use.

[0032] Next, add 500ml of water for injection into a >1000ml container, then add 35.83g of lactobionic acid in s...

Embodiment 2

[0042] The composition of each 1000ml islet isolation and purification liquid is shown in Table 2, and the balance is water for injection.

[0043] Table 2

[0044]

[0045] Specific preparation method:

[0046] First, 0.3g of CaCl 2 2H 2 O dissolved in 20ml of water for injection; 1.23g of MgSO 4 ·7H 2 O was dissolved in 10ml of water for injection; 28.06g of L-histidine was dissolved in 200ml of water for injection; 20g of polyethylene glycol-20 was heated and dissolved in 200ml of water for injection, ready for use.

[0047] Next, add 500ml of water for injection into a >1000ml container, then add 39.41g of lactobionic acid in sequence, and stir until the color of the liquid becomes clear. Add 200ml of the above-mentioned pre-made L-histidine solution, and stir until the color of the liquid becomes clear. Add 17.84g of raffinose, and stir until the color of the liquid becomes clear. Add 4.6g NaOH and stir until the liquid color becomes clear. Add 0.39g KOH and st...

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Abstract

The invention belongs to the field of biology and medicine, and specifically relates to a separating and purifying liquid for the separation and purification of pancreatic islet during the pancreatic islet transplantation and a preparation method thereof. Each 1000 mL of separating and purifying liquid contains the following components by weight: 30 to 40 grams of lactobionic acid, 4.0 to 30.0 grams of L-histidine, 15 to 20 grams of raffinose, 4.0 to 5.0 grams of NaOH, 0.35 to 0.45 gram of KOH, 1.0 to 1.5 grams of adenosine, 0.1 to 0.2 gram of allopurinol, 3.0 to 4.0 grams of NaH2PO4, 1.0 to 1.5 grams of MgSO4.7H2O, 0.05 to 0.1 gram of CaCl2.2H2O, 0.5 to 1.0 gram of glutathione, 20 to 30 grams of polyethylene glycol-20, 100,000 to 150,000 units of ulinastatin, and the balance being injection water. The pancreatic islet gradient density separating liquid can be applied to separation and purification of pancreatic islet of humans or animals, and has the advantages of better separating efficiency and protective effect, prominently lower price, and more valuable clinical application value, compared to the similar organ preservation liquid.

Description

technical field [0001] The invention belongs to the fields of biology and medicine, and specifically relates to a separation and purification liquid used in the separation and purification of islets in pancreatic islet transplantation and a preparation method thereof. Background technique [0002] Islet transplantation has become one of the ideal ways to treat type I diabetes. Whether a large number of high-purity and highly active islets can be isolated is the first prerequisite for successful islet transplantation. The islet purification method widely used in clinical practice and experiments is the gradient density centrifugation method, which uses the density difference between islets and glandular cells to purify islets. However, this separation method will lose a large number of islets, and the toxicity of the gradient separation medium will damage the activity and function of the islets, which will eventually reduce the transplantation effect. Therefore, finding a g...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 刘宝林李睿曹献馗
Owner SHENGJING HOSPITAL OF CHINA MEDICAL UNIV