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Cell culture method for inducing dental pulp stem cells to differentiate into bladder smooth muscle cells in vitro

A technology of smooth muscle cells and dental pulp stem cells, applied in the medical field, can solve the problems of limited application and poor in vitro proliferation ability of ADSCs

Inactive Publication Date: 2020-06-12
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limitations of BMMSCs and ADSCs, for example, the extraction of BMMSCs requires traumatic surgery, and the poor proliferation ability of ADSCs in vitro limits their application in smooth muscle tissue regeneration.

Method used

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  • Cell culture method for inducing dental pulp stem cells to differentiate into bladder smooth muscle cells in vitro
  • Cell culture method for inducing dental pulp stem cells to differentiate into bladder smooth muscle cells in vitro
  • Cell culture method for inducing dental pulp stem cells to differentiate into bladder smooth muscle cells in vitro

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Embodiment 1

[0034] A cell culture method for inducing dental pulp stem cells to differentiate into bladder smooth muscle cells in vitro, the cell culture method comprising the steps of:

[0035] Step (1) Isolation and cultivation of dental pulp stem cells DPSCs;

[0036] Step (2) separation and cultivation of bladder smooth muscle cells SMCs;

[0037] Step (3) collecting bladder smooth muscle cell SMCs conditioned medium CM;

[0038] Step (4) screening bladder smooth muscle cell SMCs in vitro induction culture medium;

[0039] Step (5) inducing the differentiation of dental pulp stem cells DPSCs to bladder smooth muscle cells SMCs in vitro.

Embodiment 2

[0041] Such as figure 1 As shown, a cell culture method for inducing dental pulp stem cells to differentiate into bladder smooth muscle cells in vitro, the cell culture method includes the following steps:

[0042] Step (1) Isolation and cultivation of dental pulp stem cells DPSCs;

[0043] The step (1) the separation and cultivation of dental pulp stem cells DPSCs comprises the following steps:

[0044] 1) Under sterile conditions, take out the pulp tissue from the exfoliated tooth, put the pulp tissue in an EP tube, add 1ml of collagenase type Ⅰ with a concentration of 4μg / μl, and transfer the EP tube to CO 2 Digestion was carried out in the incubator, and the EP tube was fully shaken every 20 minutes to make the pulp tissue fully contact with the digestive enzymes; after 1 hour of full digestion, the contents of the EP tube were passed through a 70 μm cell sieve and transferred to a new EP tube. Obtain a single cell suspension; use PBS buffer to fully rinse the cell suspe...

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Abstract

The invention relates to the field of cell culture, in particular to a cell culture method for inducing dental pulp stem cells to differentiate into bladder smooth muscle cells in vitro. The cell culture method comprises the following steps: (1) separating and culturing dental pulp stem cells DPSCs; (2) separating and culturing bladder smooth muscle cells SMCs; (3) collecting a bladder smooth muscle cell SMCs condition culture solution CM; (4) screening a bladder smooth muscle cell SMCs in-vitro induction culture solution; and (5) inducing the dental pulp stem cells DPSCs to differentiate intothe bladder smooth muscle cells SMCs in vitro. The differentiation of dental pulp stem cells into bladder smooth muscle cells is induced in vitro, so that the bladder smooth muscle cells can effectively replace bladder tissues of a patient needing bladder augmentation cystoplasty or replacement to play a normal role, and the patient is expected to have better living quality.

Description

technical field [0001] The invention relates to a cell culture method for applying dental pulp stem cells to bladder smooth muscle tissue engineering regeneration, in particular to a cell culture method for inducing differentiation of dental pulp stem cells into bladder smooth muscle cells in vitro. Belongs to the field of medicine. Background technique [0002] The normal detrusor muscle of the bladder expands to store urine at low pressure and contracts to urinate. Various diseases of the urinary system (including cancer, spinal injury, benign bladder spasm, etc.) will cause functional lesions of the detrusor muscle of the bladder, and such patients need to undergo cystoplasty or replacement. However, so far there is no suitable substitute that can effectively restore the normal function of the bladder detrusor. The currently commonly used cystoplasty or replacement surgery is to intercept a section of small intestine tissue, which is transplanted into the bladder after ...

Claims

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Application Information

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IPC IPC(8): C12N5/077
CPCC12N5/0661C12N2506/1353
Inventor 蒋文凯王迪雅仇珺孙书恺张亚庆宋冰余擎王胜朝
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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