Method for inducing and differentiating into dopaminergic neuron from human amniotic sepithelial cell and application for separating obtained dopaminergic neuron

An epithelial cell, dopaminergic technology, applied in the field of dopaminergic neurons to treat Parkinson's disease, can solve the problem of not finding tissue and so on

Inactive Publication Date: 2010-09-08
CHINA JAPAN FRIENDSHIP HOSPITAL
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  • Method for inducing and differentiating into dopaminergic neuron from human amniotic sepithelial cell and application for separating obtained dopaminergic neuron
  • Method for inducing and differentiating into dopaminergic neuron from human amniotic sepithelial cell and application for separating obtained dopaminergic neuron
  • Method for inducing and differentiating into dopaminergic neuron from human amniotic sepithelial cell and application for separating obtained dopaminergic neuron

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[0028] A method of inducing and differentiating dopaminergic neurons from human amniotic epithelial cells of the present invention, the method comprises the following steps:

[0029] (1) Take fresh human placenta tissue, peel off human amniotic membrane tissue, rinse repeatedly with normal saline to remove blood clots, then soak in commercially available sterile D-hank's solution for 1.5 hours, then rinse with 0.9% normal saline 3 times, sterile D-hank's solution containing: 1000U / ml penicillin, 1000U / ml streptomycin, 100U / ml gentamicin and 2.5ng / ml amphotericin;

[0030] (2) Place the rinsed human amniotic membrane in 0.25% trypsin solution, and digest it at 37° C. for 15 minutes;

[0031] (3) Use DMEM / F12 (commercially available) containing 10% fetal bovine serum to terminate the digestion of the above-mentioned human amniotic membrane in trypsin solution, then filter the above-mentioned solution with a 200-mesh cell sieve, and collect the cells obtained as human amniotic me...

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Abstract

The invention relates to a method for inducing and differentiating into dopaminergic neurons from human amniotic sepithelial cells, comprising the following steps of: taking a fresh human placenta; stripping a human amnion, repeatedly washing by using normal saline to remove blood clots, then soaking by using asepsis D-hank's liquor, and then rinsing by using the normal saline; placing the human amnion into 0.25 percent of a trypsin solution, and digesting at the temperature of 36.5-37.5 DEG C; stopping the digestion of the human amnion in the trypsin solution by using DMEM / F12 (Dulbecco's Modified Eagle Medium / F12) containing 10 percent of fetal calf serums, and then filtering the solution by using a cell sieve, wherein collected cells are the human amniotic sepithelial cells; separating the obtained human amniotic sepithelial cells, and carrying out in-vitro subculture on the separated human amniotic sepithelial cells by using 10 percent of culture mediums without the fetal calf serums; selecting 3 or 4 or 5 or 6 generation human amniotic sepithelial cells with good growth state, and then continuously culturing by using a neural stem cell differential medium; and harvesting dopaminergic neuron like cells.

Description

technical field [0001] The invention relates to a method for inducing and differentiating dopaminergic neurons from human amniotic epithelial cells and the use of the dopaminergic neurons obtained by the method to treat Parkinson's syndrome. Background technique [0002] Parkinson's disease is a spontaneous, slowly progressive, degenerative disorder of the central nervous system characterized by slowness of movement, muscle stiffness, resting tremor, and postural instability. It has now been found that it is a symptom of the progressive deterioration of neurons pigmented in the melana, locus coeruleus, and other dopaminergic cells of the brainstem, resulting in a loss of the neurotransmitter dopamine. In people over middle age, Parkinson's disease is the fourth most common neurodegenerative disease that affects people's health, affecting 0.4% of people over the age of 40 and 1% of people over the age of 65. Regardless of the patient's age, the results are often devastating ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08A61K35/30A61P25/16
Inventor 蔡哲
Owner CHINA JAPAN FRIENDSHIP HOSPITAL
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