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Pancreatic islet in-vitro culture surface parenchyma cell treatment method

A technology of in vitro culture and treatment method, applied in pancreatic cells, culture process, tissue culture, etc., can solve the problems of time-consuming and laborious operation, unsuitable for large-scale in vitro culture of pancreatic islets, etc., and achieve the effect of reducing experimental steps and reducing cell damage.

Pending Publication Date: 2021-08-27
立沃生物科技(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Multi-transfer plate culture can reduce the proportion of miscellaneous cells, but the operation is time-consuming and laborious. It is not suitable for large-scale in vitro culture of islets, and intensive operations will also have a great adverse effect on the growth status of islets.

Method used

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  • Pancreatic islet in-vitro culture surface parenchyma cell treatment method
  • Pancreatic islet in-vitro culture surface parenchyma cell treatment method
  • Pancreatic islet in-vitro culture surface parenchyma cell treatment method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] This embodiment provides a method for treating foreign cells on the surface of pancreatic islets cultured in vitro, comprising: the following steps;

[0035] S1: Spread the laminin in the culture container, ensure that the laminin solution is evenly distributed on the surface, and then put it in a CO2 incubator for incubation. During this step, the laminin matrix must not dry out.

[0036] S2: Select the isolated and purified islets to be cultured and spread them on the incubated laminin.

[0037] S3: add islet culture medium containing 5 ng / ml iodoacetic acid into the culture container, and culture for 20 h.

[0038] S4: After the culture, the islet medium containing 5 ng / ml iodoacetic acid was removed to obtain pure islets, and the islet medium without iodoacetic acid was added to the pure islets for culture.

[0039] The islet medium is RPMI 1640 medium with the following components added: 1.2g / L glucose, 10mmol / LHEPES, 10mg / L sodium pyruvate, 10% v / v fetal bovine ...

Embodiment 2

[0047] This embodiment provides a method for treating foreign cells on the surface of pancreatic islets cultured in vitro, which is characterized in that it includes the following steps;

[0048] SS1: Carry out TC treatment on the culture container and set it aside.

[0049] SS2: Select the isolated and purified islets to be cultured and place them in the TC-treated culture container.

[0050] SS3: Add the islet culture medium containing 5 ng / ml iodoacetic acid to the culture container in step SS2, and culture for 30 h.

[0051] SS4: After the culture, the islet medium containing 5 ng / ml iodoacetic acid was removed to obtain pure islets, and the islet medium without iodoacetic acid was added to the pure islets for culture.

[0052] The islet medium is RPMI 1640 medium with the following components added: 1.2g / L glucose, 10mmol / LHEPES, 10mg / L sodium pyruvate, 10% v / v fetal bovine serum, 1% v / v blue-chain Mycin; RPMI 1640 medium can be purchased commercially, such as purchased...

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Abstract

The invention discloses a pancreatic islet in-vitro culture surface parenchyma cell treatment method. The method specifically comprises the following steps: adding iodoacetic acid with a proper concentration into an islet culture medium for culturing islet, and culturing for a period of time. The strong destructive effect of iodoacetic acid on cell membranes and the inhibition capability of iodoacetic acid on cell glycolysis are utilized, and hybrid cells (especially fibroblasts) on the surfaces of pancreas islets are treated by adjusting the concentration of iodoacetic acid in the culture medium and controlling the treatment time, so that subsequent experiment steps are reduced, and cell damage is reduced.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for treating heterogeneous cells on the surface of pancreatic islet cultured in vitro. Background technique [0002] Islets are clusters of endocrine cells distributed between the acini of the exocrine part of the pancreas. The surface of pancreatic islets is coated with a thin layer of reticular fibers, but the coating is not necessarily complete in different species. Islets can be obtained by thermal digestion of pancreas with collagenase combined with density gradient centrifugation purification. That is to infuse an appropriate concentration of collagenase solution into the pancreas through the pancreatic duct, then remove the perfused pancreas and soak it in the collagenase solution, and incubate in a water bath at 37°C for a certain period of time to digest the pancreas into fine sand by collagenase and terminate the digestion of collagenase After the effect...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0676C12N2500/05
Inventor 彭园征王慈慈周明欧映廷
Owner 立沃生物科技(深圳)有限公司
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