A method for detecting the state of cell culture on a hydrophilic support
A hydrophilic scaffold and cell technology, applied in the field of cell biology detection, can solve the problems of fluorescence staining solution interference, inaccurate detection results, etc., and achieve accurate and reliable detection results, accurate detection, and facilitate large-scale promotion and application
Active Publication Date: 2019-03-01
PEKING UNIV THIRD HOSPITAL
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[0006] In order to solve the problem in the prior art that the detection of the cell culture state on the hydrophilic support is easily interfered by the fluorescent staining solution, resulting in inaccurate detection results, the embodiment of the present invention provides a method for accurately and reliably detecting the cell culture state on the hydrophilic support. state method
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Embodiment 1
[0042] This embodiment provides a method for detecting the state of cell culture on the konjac glucomannan microsphere support, comprising the following steps:
[0043] Step 1. Inoculate SD rat bone marrow mesenchymal stem cells on the konjac glucomannan microsphere scaffold for culture
[0044] Cultivating SD rat bone marrow mesenchymal stem cells: Take a male SD rat weighing 100 g, anesthetize it with an excess of urethane solution with a mass concentration of 20%, and kill it. Separate the long backbone of the rat on the sterile operating table, remove its surrounding muscle tissue and bilateral joint metaphyses, and use a 10ml syringe to extract PBS buffer (Phosphate Buffered Saline, phosphate buffered saline) to rinse the long backbone of SD rats The bone marrow cavity was washed into a 10cm culture dish to obtain a rat bone marrow mesenchymal stem cell solution. Gently pipette the rat bone marrow mesenchymal stem cell solution with a pipette to obtain a single cell susp...
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Abstract
The invention discloses a method for detecting culture state of cells on a hydrophilic scaffold and belongs to the field of cell biology detection. The method includes: inoculating to-be-detected cells onto the hydrophilic scaffold for culture, and obtaining a scaffold-cell composite system when growth fusion rate of the to-be-detected cells on the hydrophilic scaffold is 90-95%; adopting acridine orange staining liquid for fluorescent staining of the scaffold-cell composite system, and adopting a phosphate buffer solution to wash the stained scaffold-cell composite system; placing the washed scaffold-cell composite system below a laser scanning confocal microscope, and utilizing a green fluorescent passage and an orange fluorescent passage for observation to acquire the culture state of the cells on the hydrophilic scaffold. By the method, distribution condition and growth state of the cells on the hydrophilic scaffold can be detected clearly and accurately; the method is simple and convenient to operate and convenient for large-scale popularization and application.
Description
technical field [0001] The invention relates to the field of cell biology detection, in particular to a method for detecting the state of cell culture on a hydrophilic support. Background technique [0002] Tissue engineering scaffold is a biological substitute for repairing, maintaining, and promoting the functions and shapes of various tissues or organs after injury. It can deliver nutrients and excrete metabolites for cell growth, and provide support for cell proliferation and differentiation. Provide structural support and space. In order to know the effect of tissue engineering scaffolds, when using tissue engineering scaffolds for cell culture, it is necessary to detect the distribution, growth and proliferation of cultured cells on tissue engineering scaffolds. Therefore, it is necessary to provide a simple, clear and accurate method for detecting the state of cell culture on tissue engineering scaffolds. [0003] At present, the existing technology usually uses flu...
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IPC IPC(8): C12Q1/02
Inventor 敖英芳孟庆阳黄洪杰皮彦斌袁兰
Owner PEKING UNIV THIRD HOSPITAL