Mesenchymal cell marking and tracing method
A cell and labeling technology, applied in the field of medical cell engineering, can solve the problems of low efficiency, low accuracy, long time consumption, etc., and achieve the effects of accurate detection, simple method and small radiation effect.
Inactive Publication Date: 2019-12-03
孙桂珍
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[0003] The current traditional labeling and tracing methods for mesenchymal cells are inefficient, time-consuming, and less accurate. In view of the above, here we propose a method for labeling and tracing mesenchymal cells
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[0016] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Apparently, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.
[0017] The present invention provides a technical solution: a method for labeling and tracing mesenchymal cells, including the following method steps, S1, transferring mesenchymal cells containing a target gene into a nutrient medium, and inducing the cells to carry out multiple Split;
[0018] S2, then add glycerol or dimethyl sulfoxide protective agent to the culture medium, place in liquid nitrogen, and control the osmotic pressure to 250-325mmol / L;
[0019] S3, then cut out the mesenc...
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The invention discloses a mesenchymal cell marking and tracing method. The method comprises the following steps: S1, transferring a mesenchymal cell containing a target gene into a nutrient medium andinducing the cell to carry out cell divisionmultiple times; S2, adding glycerol or a dimethyl sulfoxide protective agent into a culture solution, putting the mixed culture solution into liquid nitrogen, and controlling the osmotic pressure to be 250 to 325mmol / L; S3, cutting the mesenchymal cell, putting the cut mesenchymal cell into 100% of glucose for 10 seconds, carrying out full immersion andenabling a membrane to be semitransparent, and carrying out shaking for 3 minutes; S4, carrying out full marking by using fluorescent dye; and S5, carrying out tracing by usingan isotope tracing method. According to the method, various fluorescent dye materials are used for marking; the overall sensitivity is high; the method is simple and convenient; the experiment process is simplified; positioning and quantification are accurate; and physiological conditions are met. Meanwhile, the isotope tracing method can be used for rapid positioning; the radiation effect is small; detection is accurate; and the whole method is scientific, efficient and is worthy of popularization.
Description
technical field [0001] The invention relates to the technical field of medical cell engineering, in particular to a method for marking and tracing mesenchymal cells. Background technique [0002] Mesenchymal cells are poorly differentiated cells with large nuclei, oval eggs, prominent nucleoli, and weakly basophilic cytoplasm. In addition to a certain number of mitochondria in the cytoplasm, there are also a small amount of rough endoplasmic reticulum, free ribosomes, Golgi apparatus, scattered lysosomes and liposomes, etc. The interstitium of the mesenchyme is a colorless and transparent liquid. In addition to a certain number of mitochondria, there are also a small amount of rough endoplasmic reticulum, free ribosomes, Golgi bodies, scattered lysosomes and liposomes in the cytoplasm. . The mesenchyme is a colorless, transparent liquid. It has a strong ability to divide and differentiate. In the process of embryonic development, it is not only the common ancestor of vari...
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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6486
Inventor 孙桂珍
Owner 孙桂珍