Cryopreservation method for dental organ
A cryopreservation method and organoid technology, applied in the biological field, can solve the problems of immaturity, loss, and weak developmental ability of autologous tooth transplantation and tooth replantation, and achieve the effect of eliminating time dependence.
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Embodiment 1
[0058] A dental organ cryopreservation method, the specific steps are as follows:
[0059] (1) Obtaining donor dental organoids; dental organoids can be tooth germs.
[0060] (2) Dental organoids are soaked in a container with medium, let stand for 8 minutes, and then store the container under freezing conditions for 12 hours and then transfer to liquid nitrogen for storage; the medium contains 8% dimethylformazan Sulfoxide-based BGJb medium; the freezing temperature is -70°C; the concentration of liquid nitrogen is ≥98%.
[0061] (3) Place the container in a 37°C water bath for rapid thawing and recovery when in use;
[0062] (4) Take out the dental organoids, wash them with PBS buffer solution, and soak them in PBS buffer solution or normal saline until ready to use. Wash can be washed 2 times.
[0063] The container was stored at -80°C for 24 hours; said; the mass percent concentration of dimethyl sulfoxide was 10%.
[0064] The way to obtain donor dental organoids is t...
Embodiment 2
[0068] A dental organ cryopreservation method, the specific steps are as follows:
[0069] (1) Obtain donor dental organoids; dental organoids can be maxillary first molar tooth germs. Use the tooth germs that have not yet started to develop and have no hard tissue at the root for cryopreservation, and try to recycle the tooth germs. Tooth germs can still develop physiological roots after cryopreservation. The cryoprotective solution can penetrate into the hard tissue-wrapped dental papilla, so that the dental papilla retains good activity after resuscitation and continues to develop into a pulp-dentin complex.
[0070] (2) Dental organoids were soaked in a container containing a culture medium, left to stand for 12 minutes, and then stored in a freezer for 36 hours, then transferred to liquid nitrogen for storage; the culture medium was 12% dimethyl sulfoxide The BGJb medium; the freezing temperature is -75°C; the liquid nitrogen concentration is ≥98%.
[0071] (3) Place t...
Embodiment 3
[0078] A dental organ cryopreservation method, the specific steps are as follows:
[0079] (1) Obtain donor dental organoids; dental organoids can be maxillary first molar tooth germs, use tooth germs whose roots have not yet begun to develop, and have no hard tissues at the roots for cryopreservation, and try to recycle tooth germs.
[0080] On the eighth day after birth, the tooth germ of the maxillary first molar of the mouse is in the bell-shaped terminal stage, and the root is soft tissue, which is conducive to the liquid exchange of the culture medium. In the present invention, tooth germs can still develop physiological tooth roots after cryopreservation, and tooth germs can be collected and preserved as a resource, and can also provide an experimental basis for the construction of tissue-engineered teeth or in vitro organoids in the future.
[0081] (2) Dental organoids were soaked in a container containing a culture medium, left to stand for 10 minutes, and then the c...
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