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Primary culture method for pancreas acinar cells

A technology of acinar cells and primary culture, applied in the field of primary culture of pancreatic acinar cells, can solve problems such as difficult culture, and achieve the effect of high difficulty and high purity

Inactive Publication Date: 2012-10-03
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a stable primary culture method for pancreatic acinar cells that can be successfully passed down according to the difficulty of in vitro primary culture in the existing culture of pancreatic acinar cells

Method used

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  • Primary culture method for pancreas acinar cells
  • Primary culture method for pancreas acinar cells
  • Primary culture method for pancreas acinar cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Separation and Purification of Pancreatic Acinar Cells

[0029] 1. Materials and Methods

[0030] 1.1 Animals

[0031] Healthy, clean-grade SD rats were fed with common feed, male or female, weighing 250-300 g (provided by Guangdong Medical Experimental Animal Center).

[0032] 1.2 Reagents

[0033] D-Hanks Solution; Percoll Cell Separation Solution (Pharmacia); Normal Saline; Digestion Solution: Type I Collagenase (Sigma), when used, prepare 0.5g / L cells with freshly prepared D-Hanks solution (without calcium ions) Separation fluid; 100ml / L calf serum; DMEM / F12 dry powder medium (Gibico). Other ingredients: penicillin (Penicillin); streptomycin (Streptomycin); amphotericin B (AmphotericinB); soybean trypsin inhibitor (Soybean Trypsin Inhibitor, SBTI); epidermal growth factor (Epidermal Growth Factor, EGF); Beijing Dingguo); pancreatic acinar antibody; glutaraldehyde; paraformaldehyde; osmic acid; PBS; ethanol; embedding agent; uranyl acetate; acetone; 4wt...

Embodiment 2

[0041] Example 2 Identification of pancreatic acinar cells

[0042] 1 Morphological observation of pancreatic acinar cells

[0043] Before and after the purification of pancreatic acinar cells, the pancreatic acinar cells cultured for 12h, 1d, 3d, 5d, 9d, 15d, and 30d were all observed and photographed under a light microscope.

[0044] Freshly isolated rat pancreatic acinar cells were round or oval under the light microscope, mixed with irregular interstitial cells and cell debris ( figure 1 ). After the cells were separated and purified by Percoll cell separation medium, most of the acinar cells were found to be round or oval in suspension, the ratio of cytoplasm to nucleus was large, the nucleus was round and biased to one side, and there were few intercellular impurities ( figure 2 ). After 12 hours, the cells can be seen to aggregate into a string of beads, and individual cells become larger, elongated, and in a state of division. ( image 3 ). After 24 hours, the ...

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Abstract

The invention discloses a primary culture method for pancreas acinar cells. The primary culture method for the pancreas acinar cells comprises the following steps of: (1) preparing pancreas acinar cell suspension; and (2) separating and purifying the pancreas acinar cells by a Percoll discontinuous density gradient precipitation method. The method can successfully culture the pancreas acinar cells, smoothly perform passage and lay a foundation for pathogenesis or pharmacotherapy of related diseases of the pancreas acinar cells; and the activity of the cells reaches over 96 percent.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for primary culture of pancreatic acinar cells. Background technique [0002] Although there is a certain understanding and understanding of the pathophysiological process of pancreatic disease, its exact pathogenesis is still unclear, especially the curative effect of severe acute pancreatitis is still not satisfactory. Studies have found that acute pancreatitis begins in acinar cells, but in clinical practice, the difficulty in collecting pancreatic specimens has seriously affected the conduct of clinical trials. In the laboratory, since pancreatic acinar cells are cells that can secrete pancreatic enzymes and digest themselves, this increases the difficulty of primary culture of acinar cells in vitro. At present, there are few literatures about the primary culture of pancreatic acinar cells published abroad, and the longest culture time does not exceed 20 days, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
Inventor 陈恳崔淑兰陈婧华龙友明王晖葛全兴
Owner GUANGDONG PHARMA UNIV
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