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Formula of density gradient solution used for tissue and cell purification

A technology of density gradient and tissue, which is applied in the field of density gradient solution formulation, can solve the problems such as not being suitable for the islet separation center, achieve ideal tissue and cell separation results, be convenient to use, and promote the effect of cell transplantation research and application

Inactive Publication Date: 2016-08-24
WENZHOU YIKANG CELL TRANSPLANTATION CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this adjustment may not be suitable for all islet isolation centers, because the optimal use temperature of UW solution is at 4°C, so in order to ensure the viability of islets, COBE 2991 must be modified and run at this temperature, placed in a cold Room

Method used

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  • Formula of density gradient solution used for tissue and cell purification
  • Formula of density gradient solution used for tissue and cell purification
  • Formula of density gradient solution used for tissue and cell purification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The formula of the density gradient liquid used in a tissue and cell purification process in Example 1 of the present invention is as follows:

[0039] (1) Formula 1: Iodixanol 248.0g, potassium lactobionate 15g, gadopentetate meglumine 5g, RPMI1640 to 1L.

[0040] (2) Preparation process:

[0041] 1) Take the components of RPMI 1640 (the components of RPMI 1640 can refer to the following table 1), first dissolve with 80% three-distilled water, if necessary, first use HCl to adjust the acid to dissolve, and then use NaOH to adjust the pH value to neutralize, Dilute to 1L with triple distilled water.

[0042] Table 1 RPMI 1640 Component List

[0043]

[0044]

[0045] 2) Accurately weigh 248.0g of iodixanol, 15g of potassium lactobionate, and 5g of gadopentetate meglumine, put them into a 1L sterilized glass bottle, dissolve them in 800ml of the above-mentioned RPMI 1640 solution, and then use the above-mentioned RPMI 1640 solution to dilute to 1L. If necessary...

Embodiment 2

[0064] The formula of high-density density gradient liquid in a tissue and cell purification process of Example 2 of the present invention is as follows:

[0065] (1) Formula: iodixanol 204.0g, diatrizoate sodium 7.36g, gadopentetate meglumine 10.0g, CMRL 1066 to 1L.

[0066] (2) Preparation process:

[0067] 1) Take the components of CMRL 1066 (the components of CMRL 1066 can refer to the following table 2), and first dissolve them with 80% three-distilled water, if necessary, first use HCl to adjust the acid to dissolve, and then use NaOH to adjust the pH value to neutralize, Dilute to 1L with triple distilled water.

[0068] Table 2 CMRL 1066 ingredient list

[0069]

[0070]

[0071] 2) Accurately weigh 204.0g of iodixanol, 7.36g of sodium diatrizoate, and 10.0g of gadopentetate meglumine into a 1L sterilized glass bottle, dissolve with 800ml of the above CMRL 1066 solution, and then use the above CMRL 1066 solution Dilute to 1L. If necessary, use 1N HCl or NaOH ...

Embodiment 3

[0077] A low-density density gradient liquid formula in the tissue and cell purification process of Example 3 of the present invention is as follows:

[0078] (1) Formula: iodixanol 10.22g, diatrizoate sodium 3.59g, gadopentetate meglumine 15g, CMRL 1066 to 250ml.

[0079] (2) Preparation process:

[0080] 1) Accurately weigh 10.22g of iodixanol, 3.59g of sodium diatrizoate, and 15g of gadopentetate meglumine into a 1L sterilized glass bottle, dissolve with 200ml of the above CMRL 1066 solution, and then use the above CMRL 1066 solution to make up to volume to 250ml. If necessary, use 1N HCl or NaOH to adjust the pH to 7.2±0.2, and measure the density with a hand-held electronic density meter to be 1.06±0.01g / ml. If not, add a small amount of iodixanol, sodium diatrizoate or CMRL 1066 to check the density Adjustment.

[0081] 3) In the local A-level clean area under the B-level background, first filter with 0.45 μm microporous membrane and then with 0.22 μm microporous memb...

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Abstract

The invention provides a formula of a density gradient solution used for tissue and cell purification. According to the formula, the density gradient solution contains density gradient ingredients, such as iohexol, sodium diatrizoate and iodixanol, wherein the substances capable of improving the osmotic pressure and preventing tissue and cell edema, such as raffinose and mycose are added, then magnevist solution is also added, and culture solutions such as DMEM culture solution and buffer solutions such as HBSS buffer solution are adopted as the solvents. With the adoption of the formula, the problems that at present, the density gradient separation for tissues and cells of large animals and human beings is difficult, and the separated tissues and cells are low in activity are solved, and the cells and tissues with different densities after dissociation or mechanical dispersion can be separated by the density gradient solution under the centrifugal condition. Due to the design of the density gradient solution, a certain application value is achieved for promoting the domestic and overseas study on the separation of tissues and cells. The principle is simple, the preparation is easy, the use is convenient, and the large-scale preparation can be realized; due to the declaration of the patent, a foundation is laid for the development and application of the density gradient separating solution.

Description

technical field [0001] The invention relates to a density gradient solution formula used alone or in combination with other solutions in the process of tissue and cell purification, in particular to a density gradient solution for tissue and cell purification based on cell culture fluid, which is used for animal Or density gradient purification of human tissues and cells. Background technique [0002] The famous American biologist George Daly said: "The 20th century is the era of drug therapy, and the 21st century is the era of cell therapy." The development of human transplantation is one of the most outstanding achievements in medicine in the 20th century. Especially from the late 1970s to the early 1980s, new immunosuppressants came out, which greatly improved the survival rate of grafts and the clinical efficacy of organ transplantation. improve. Due to the large increase in clinical transplant cases in recent years, the shortage of donors is very prominent. In the 21...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0602C12N5/0676C12N2509/00
Inventor 傅红兴刘成洋蒋煊赵应征
Owner WENZHOU YIKANG CELL TRANSPLANTATION CO LTD
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