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Separation and culture method of gingival stem cells and application of gingival stem cells

A stem cell and gingival technology, applied in the field of stem cell culture, can solve problems such as the inability of the aqueous humor to flow smoothly, the failure of glaucoma surgery, and the blockage of the aqueous filtration tract, etc., so as to shorten the digestion processing time, delay the generation of non-functional filtering bubbles, increase the effect of time

Pending Publication Date: 2020-12-04
北京泰生生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, scar formation after anti-glaucoma surgery blocks the smooth filtration channel of aqueous humor, resulting in unsmooth flow of aqueous humor, intraocular pressure rises again, and failure of glaucoma surgery

Method used

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  • Separation and culture method of gingival stem cells and application of gingival stem cells
  • Separation and culture method of gingival stem cells and application of gingival stem cells
  • Separation and culture method of gingival stem cells and application of gingival stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Isolation and cultivation of gingival stem cells

[0052] 1. Pretreatment of gingival tissue

[0053] 1-1) Put the teeth (primary teeth or third molars) of healthy subjects removed from the operating table into a collection solution containing 3% fetal bovine serum and penicillin-streptomycin (10,000 U / ml) of phosphate buffer solution, stored at low temperature, and transferred to the laboratory; then the collected teeth were poured into 100mm Petri dishes, and the surface of the crown was disinfected with iodophor cotton swabs, and the crown was clamped with a needle holder, Use a disposable dropper to suck the cleaning solution to clean the tooth surface, wherein the cleaning solution is a phosphate buffer solution containing 1 volume of penicillin and streptomycin (10,000 U / ml);

[0054] The phosphate buffer is PBS commonly used in the art, that is, neutral phosphate buffer solution. The penicillin and streptomycin in the collection solution and the clea...

Embodiment 2

[0068] The identification of embodiment 2 gingival stem cells

[0069] The gingival stem cells used in the following experiments are the second to fifth passage gingival stem cells cultured according to the above method.

[0070] The methods for cell immunophenotype identification and adipogenic and osteogenic differentiation identification of isolated and cultured gingival stem cells in the present invention refer to the reference method for cell phenotype identification and adipogenic and osteogenic differentiation identification of oral cavity-related stem cells (GANG DING, WEIWANG, YI LIU, et al.Effect of Cryopreservation on Biological and ImmunologicalProperties of Stem Cells From Apical Papilla.JOURNAL OF CELLULAR PHYSIOLOGY,J.Cell.Physiol.223:415–422,2010.);(F Feng1,K Akiyama2,Y Liu , et al, Utility of PDL progenitors for in vivo tissue regeneration: a report of 3cases. Oral Diseases (2010) 16, 20–28.); (Takayoshi Yamaza, Akiyama Kentaro, Chider Chen, et al. Immunomodul...

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Abstract

The invention discloses a separation and culture method of gingival stem cells and application of the gingival stem cells. The separation and culture method comprises the step of carrying out digestion treatment on gingival tissues by adopting a collagenase-phosphate buffer solution with the mass volume concentration of less than 0.3%. According to the separation method disclosed by the invention,the gingival tissues are subjected to the digestion treatment by only adopting I-type collagenase, so that the quantity of obtained primary gingival stem cells is great and the survival rate is high;the separation speed is rapid and the treatment time is short, so that the cost of preparing the stem cells is remarkably reduced; after a glaucoma operation, the gingival stem cells separated and cultured by the method can be used for remarkably prolonging the time of transition of filter bubbles from functional filter bubbles to non-functional filter bubbles and keeping an aqueous humor filtering channel smooth, so that aqueous humor can be smooth and the intraocular pressure is prevented from being raised again after the operation; and the gingival stem cells have the effect of enhancing afiltering function after an anti-glaucoma filtering operation and the success rate of the glaucoma operation is improved.

Description

technical field [0001] The invention relates to a method for primary isolation and culture of stem cells, in particular to a method for separation and culture of gingival stem cells and the application of gingival stem cells, belonging to the field of stem cell culture. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of pluripotent stem cells belonging to the mesoderm. They mainly exist in connective tissue and organ interstitium, and are most abundant in bone marrow tissue. Since bone marrow is its main source, they are collectively called bone marrow mesenchymal cells. stem cells. Mesenchymal stem cells have all the common characteristics of stem cells, that is, self-renewal and multi-directional differentiation capabilities, directional migration to tissue damage sites, promotion of tissue repair, nerve protection and immune regulation. At present, it is also the most clinically used, and combined with hematopoietic stem cells can improve the succe...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28A61P27/06
CPCC12N5/0668A61K35/28A61P27/06C12N2509/00
Inventor 不公告发明人
Owner 北京泰生生物科技有限公司
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