Isolation and culture method of human adipose-derived stem cells and construction method of stem cell bank
A technology for the isolation and cultivation of human adipose stem cells, which is applied in the construction of stem cell banks and the field of isolation and cultivation of human adipose stem cells, can solve the problems of slow growth of human adipose stem cells, severe cell differentiation, and unsatisfactory conditions, and achieve short culture time and stem cell The effect of high purity and fast growth
Inactive Publication Date: 2019-06-07
CENTURY BIOSTRENGTH BEIJING PTY LTD
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Problems solved by technology
[0003] At present, in the isolation and culture of human adipose stem cells, the growth rate of human adipose stem cells is slow, and the cell differentiation is serious, which cannot meet the needs of clinical research and future medical use
Method used
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Embodiment 1
[0045] A culture medium for human adipose stem cells, DMEM-F12 medium with a final concentration of 1% fetal bovine serum, 1ng / ml of bFGF and 1ng / ml of EGF.
Embodiment 2
[0047] A culture medium for human adipose stem cells, in which DMEM-F12 medium is supplemented with fetal bovine serum at a final concentration of 20%, bFGF of 100 ng / ml and EGF of 100 ng / ml.
Embodiment 3
[0049] A culture medium for human adipose stem cells, DMEM-F12 medium is supplemented with fetal bovine serum at a final concentration of 10%, bFGF at 10 ng / ml and EGF at 10 ng / ml.
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The invention discloses an isolation and culture method of human adipose-derived stem cells. A culture medium is a DMEM-F12 medium added with fetal calf serum with final concentration of 1-20%, bFGF of 1-100 ng / ml and EGF of 1-100 ng / ml. The isolation and culture method includes fat primary operation steps. The fat primary operation steps include steps of crushing, cleaning, digestion, isolation and culturing. The isolation and culture method also includes subculture operation steps. The subculture operation steps include cleaning, digestion, cleaning, centrifugation and culturing. The isolation and culture method of the human adipose-derived stem cells is simple in culture medium composition, short in culture time in the isolation and culture method, fast in growth speed and high in stemcell purity. No stem cell differentiation occurs during the culture.
Description
technical field [0001] The invention belongs to the field of stem cells, and in particular relates to a method for separating and culturing human adipose stem cells and a method for constructing a stem cell bank. Background technique [0002] In 2001, the multi-lineage stem cells were first isolated from human adipose tissue suspension by liposuction. Because it is easy to extract stem cells from adipose tissue, and the adipose tissue suspension discarded in liposuction can be turned into an important source of human stem cell bank, so the research in this area has been carried out rapidly and deeply. It has been proven that adipose tissue-derived stem cells (ATSCs) can be induced and transformed into adipocytes, chondrocytes, osteoblasts, muscle cells, and cardiomyocytes. They can be used as seed cells for various tissue engineering, and have very important research and application values. [0003] In the current isolation and culture of human adipose stem cells, the growt...
Claims
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IPC IPC(8): C12N5/0775C40B50/06
Inventor 董明珠李霞云潘新刘世红卢家堃
Owner CENTURY BIOSTRENGTH BEIJING PTY LTD
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