Method for evoking differentiation of mesenchyma stem cell and fat stem cell into neurons and special culture medium thereof
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A technology of adipose stem cells and mesenchymal stem cells, which is applied in the field of cell culture methods and its special medium, and can solve problems such as toxic and side effects
Active Publication Date: 2008-02-06
FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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[0002] Neuron damage diseases (such as Parkinson's disease, Alzheimer's disease, cerebral ischemia, etc.) are major diseases that currently plague human beings. At present, there is
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Embodiment 1
[0037] Example 1. Inducing differentiation of mesenchymal stem cells into neurons
[0038] Taking mesenchymal stem cells derived from iliac bone marrow as an example, the differentiation induction effect of the method of the present invention was tested. Bone marrow mesenchymal stem cells can be obtained by the operation in the following step 1, or directly obtained from a stem cell bank.
[0039] 1. Isolation, purification, primary and subculture of bone marrow mesenchymal stem cells
[0040] Obtain iliac bone marrow from surgically discarded iliac bone marrow, or extract ilium bone marrow from healthy voluntary donors under sterile conditions, or obtain bone marrow from a bone marrow bank, add DMEM-F12 culture medium (high-glucose DMEM medium and DF-12 medium (product of Sigma Company, Cat#D0547) was obtained by mixing at a volume ratio of 1:1), centrifuged, discarded the supernatant and fat layer, and fully mixed the bone marrow cell pellet with DMEM-F12 medium Finally, g...
Embodiment 2
[0084] Example 2. Inducing the differentiation of adipose-derived stem cells into neurons
[0085] Taking adipose-derived stem cells as an example, the differentiation induction effect of the method of the present invention was tested. Adipose stem cells can be obtained by the operation in the following step 1, or directly obtained from a stem cell bank.
[0086] 1. Isolation, purification, primary and subculture of adipose-derived stem cells
[0087] 1) Use the fat discarded during liposuction or absorb fat from healthy volunteers by liposuction, rinse with PBS, add 4 times the volume of 0.1% collagenase, 1 times the volume of 2% BSA and double antibody, and then Put it into a 50mL centrifuge tube, digest at 37°C for 45min, and shake it from time to time.
[0088] 2) Filter the adipose tissue that has been digested into a paste with an 80-mesh sieve, and then filter it again with a 200-mesh sieve. Add collagenase to the remaining adipose tissue to continue digestion and rep...
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Abstract
The present invention discloses a method for inducing mesenchymal stem cell to divide into nerve cell and the special culture medium. The culture medium is DMEM culture medium of cattle fetus blood serum which the volume and the thickness percent ratio is 8 to 12 percent. The method is provided with steps as follows: firstly, a small disturbing RNA which restrains the express of limited silent gene of a nerve cell is composed or an intervention carrier of carrying a small disturbing RNA coding gene with restraining the express of limited silent gene of a nerve cell is composed; secondly, the small disturbing RNA which restrains the express of limited silent gene of a nerve cell or the intervention carrier of carrying a small disturbing RNA coding gene with restraining the express of limited silent gene of a nerve cell is transformed or transfected into mesenchymal stem cell or fat stem cell, then the recombined mesenchymal stem cell or the recombined fat stem cell is positioned in the culture medium induced by nerve cell, and cultivated under the temperature of 36 to 38 DEG C, thereby getting nerve cell. The prevent invention can play an important role in the medical field, and has wide applied prospect.
Description
technical field [0001] The invention relates to a cell culture method and a special medium thereof, in particular to a method for inducing differentiation of mesenchymal stem cells (Mesenchymal stem cells, MSCs) and adipose stem cells into neurons and a special medium thereof. Background technique [0002] Neuron damage diseases (such as Parkinson's disease, Alzheimer's disease, cerebral ischemia, etc.) are major diseases that currently plague human beings. At present, there is no drug that can cure neuron damage diseases. Some drugs used to treat the disease Symptoms can only be alleviated to some extent, and long-term use can also produce toxic side effects. In recent years, the use of stem cell transplantation to treat neuron damage diseases has received extensive attention from the medical community. Mesenchymal stem cells (MSCs) are highly expected due to their easy acquisition, low immunogenicity, strong proliferation ability, multidirectional differentiation potentia...
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