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A placenta potential cell and a culture method thereof
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A technology of potential cells and culture methods, which is applied in the field of potential cells derived from animal placenta and its culture, and can solve the problems of poor growth and proliferation of cells
Inactive Publication Date: 2017-11-17
SHANGHAI STEMSAN BIOTECH CO LTD
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When used alone, the cells survive but cannot proliferate well
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Embodiment 1
[0110] Embodiment 1 Culture human placenta cell in vitro
[0111] To obtain human placenta, first put the placental tissue into 4°C pre-cooled phosphate-buffered saline (PBS) containing double antibodies (penicillin and streptomycin), wash 3 times, and then cut the large tissue block into 2mm For small tissue pieces, wash twice with cold PBS containing double antibodies, and put the washed tissue pieces into 0.25% neutral protease solution or 1% collagenase solution prepared with sterile PBS for digestion, and the condition is constant temperature at 37°C Oscillate for 1 hour, and then carry out experiments according to the following methods.
[0112] Pipette the tissue block repeatedly with a pipette or a pipette, or pour the tissue block together with the digestive solution onto a stainless steel filter, and grind the tissue block with a syringe plunger until the tissue block is completely dispersed and becomes a single cell.
[0113] This contains cells and a digestive enz...
Embodiment 2
[0124] Example 2 Formation of Induced Differentiation of Adipose Tissue, Bone and Cartilage
[0125] see image 3 , continue to culture the placental cells of the experimental group, adding three inducers that form bone, cartilage, and fat. After two weeks, adipose tissue, bone, and cartilage are formed.
[0126] see Figure 4-5 , using different staining methods, under the microscope, to form bone, cartilage, and adipose tissue.
Embodiment 3
[0127] Example 3, identification of cell surface markers
[0128] Referring to Fig. 7, the same method as in Example 1 was used for culturing, and a control group and an experimental group were set up, and no substance of the present invention was added to the control group. After 30 days of culture, the cell surface markers were measured by flow cytometry. see Figures 7A-7I , the experimental results are shown below. Figure 7A Show CD90>95%, Figure 7B Show CD34 Figure 7C show CD14 Figure 7D show CD45 Figure 7E show CD19 Figure 7F Show CD105>95%, Figure 7G Show CD31 Figure 7H Show CD73>95%, Figure 7I Show HLA-DR<2%. The cells in the experimental group express CD90, CD73, and CD105, but do not express CD45, CD14, and HLA-DR, and the control group does not express them.
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Abstract
A culture method of a placenta potential cell is disclosed. The method includes (1) preparing placental cells and / or tissues under sterile conditions; (2) inoculating the placental cells and / or tissues into a medium, culturing the placental cells and / or tissues, and adding a cell growth regulator into the medium to allow the placenta potential cell to grow so that the placental cells and / or tissues are brought into an amplification state; and (3) culturing the placenta potential cell to allow the placenta potential cell to be continuously proliferated into a cell having stem cell characteristics. A living matter capable of maintaining and supporting the potential cell is found, and replaces a medicine. Through tissue and organ replication, tissues and organs are physiologically repaired, the balance of vital organs is maintained, difficult, chronic and stubborn symptoms and diseases are treated and a patient is allowed to get rid of medical treatment pain and damage. Cancer is expected to be treated by utilizing an adaptive cell regulation method.
Description
technical field [0001] The invention relates to a potential cell and a culture method thereof, in particular to a potential cell derived from an animal placenta and a culture method thereof. Background technique [0002] Since the end of the 20th century, life science research has discovered that stem cells can be cultured in vitro to form cells / tissues characteristic of various organs of the human body. Therefore, in vitro culture of "constructed organs" is expected to carry out "autologous organ" transplantation to treat diseases in the near future; or, inject stem cells into a certain part of the human body, let stem cells play the function of an organ locally, and finally realize the cure for diseases. treat. However, in the process of stem cell culture, in vitro stem cell culture technology is limited by the rapid aging and self-differentiation of stem cells and the loss of stem cell stemness. [0003] In the prior art, cell culture refers to a culture technology that...
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Application Information
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