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Cryopreservation method for protecting cell junction

A technology of cell connection and preservation method, which is applied in the field of cryogenic biological preservation and can solve the problems of impermeability of cell membranes and the like

Inactive Publication Date: 2012-06-13
HOSPITAL ATTACHED TO QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, unlike dimethyl sulfoxide, ethylene glycol, etc. (permeable protective agents), trehalose is an impermeable substance that cannot penetrate the cell membrane and can only play a protective role outside the cell.

Method used

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  • Cryopreservation method for protecting cell junction
  • Cryopreservation method for protecting cell junction
  • Cryopreservation method for protecting cell junction

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0136] experiment material

[0137] The thoracic aortas of 24 200-250g SD rats were carefully cleaned of connective tissue around the blood vessels, cut into several arterial rings about 3-5mm in length, and divided into 3 groups: fresh control group and routine preservation group (conventional cryopreservation) , Experimental group (high heat shock protein expression + trehalose preservation).

[0138] 2. Main reagents

[0139] Dimethyl sulfoxide (Me 2 SO), glutamine (Glu), trehalose, cell culture medium (DMEM)

[0140] 3. Experimental method

[0141] High expression method of heat shock protein

[0142]The three groups were treated with high expression of heat shock protein: glutamine (Gln) intravenous injection of 0.75g·kg -1 , blood vessels were taken after 6 hours (n=8).

[0143] Conventional cryopreservation protectant (CPM)

[0144] No. 1 CPM: 5% (v / v) Me 2 SO+DMEM culture medium; No. 2 CPM: 10% (v / v) Me 2 SO+DMEM culture solution.

[0145] Trehalose cryoprote...

example 2

[0175] 1. Experimental materials and methods

[0176] Isolation, Culture and Osteogenic Differentiation of cBMSCs (Canine Bone Marrow Mesenchymal Stem Cells)

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Abstract

The invention discloses a cryopreservation method for protecting cell junction. The method comprises the following steps of: a) culturing Madin Darby canine kidney (MDCK) cells; b) dividing the MDCK cells in the step a) into a normal control group, a cryopreservation group, a trehalose group, an Hsp70 high expression group and an Hsp70+ trehalose group; c) performing filter culture on the normal control group, the cryopreservation group and the trehalose group respectively, performing filter culture and gene transfection on the Hsp70 high expression group, and performing filter culture and gene transfection on the Hsp70+ trehalose group; d) performing cryopreservation on the normal control group, the cryopreservation group, the trehalose group, the Hsp70 high expression group and the Hsp70+ trehalose group; e) growing cells and performing morphologic observation (the cultured cells are observed by a microscope and are subjected to HE staining); f) testing the survival rate of the cells; g) observing and analyzing the cell junction morphology and structure; and h) performing a fluorescein experiment. The method is simple, and convenient to use; and the structure and function of the cell junction after tissues and organs are subjected to cryopreservation can be kept complete, and the damage of the cell junction can be effectively avoided.

Description

Technical field: [0001] The invention relates to the technical field of cryogenic biological preservation, in particular to a cryopreservation method for protecting cell connections, which is beneficial to the preservation of tissues and organs. Background technique: [0002] The cryopreservation of tissues and organs is a complex problem plaguing the field of cryobiology research. The position, structure and function of cell junctions in tissues are special, so their response to freezing injury and the protection mechanism of freezing injury should also have their particularity. Therefore, it is necessary to systematically observe and study the characteristics and rules of cell junction damage in cryopreservation, establish targeted and effective cryopreservation technical solutions, and improve and ensure the cryopreservation effect of tissues and organs. Studies have shown that cell junctions may be one of the targets of various damage factors during cryopreservation, le...

Claims

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Application Information

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IPC IPC(8): A01N1/02
Inventor 王沛涛
Owner HOSPITAL ATTACHED TO QINGDAO UNIV
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