Method for extracting amniotic stem cells

Abstract

The invention discloses a method for extracting amniotic stem cells. The method comprises the following steps: A1, mechanically separating amniotic epidermis from a healthy placenta, and performing washing; a2, placing a first culture dish in a second culture dish, wherein a plurality of holes are formed in the bottom surface of the first culture dish; a3, cutting the cleaned amnion into pieces, putting the amnion pieces into a first culture dish, adding a buffer solution, then taking out the first culture dish, and paving cut amnion tissue blocks at the bottom of the first culture dish; and A4, putting the first culture dish with the amnion tissue block into another second culture dish again, adding the culture solution for culture, allowing standing for 15-18 days, adding pancreatin fordigestion, then adding the culture solution to stop digestion, taking out the first culture dish, and then extracting stem cells from the culture solution mixed with the cells. The method can effectively solve the problems of high cost and low efficiency in the prior art and the operation complexity caused by a manual inoculation mode, can quickly achieve extraction operation, reduces the cost andimproves the working efficiency.

Description

technical field [0001] The invention belongs to the technical field of stem cell extraction and culture, and in particular relates to a method for extracting amniotic membrane stem cells. Background technique [0002] Amniotic stem cells are stem cells isolated from the amniotic membrane of newborn fetuses, including amniotic epithelial stem cells and amniotic mesenchymal stem cells. In addition, a large number of stem cells can also be isolated from amniotic fluid. Since amniotic fluid is not easy to collect and is easily polluted, the application of amniotic fluid-derived stem cells is also limited. [0003] Advantages of amniotic stem cells [0004] (1) Amniotic membrane stem cells (including amniotic membrane epithelial stem cells and amniotic membrane mesenchymal stem cells) have abundant sources, simple isolation process, strong multi-differentiation potential, no ethical barriers, strong proliferation ability, no tumorigenic effect in vivo and in vitro, and low immun...

AI Technical Summary

Problems solved by technology

[0009] At present, there are two traditional extraction methods of amniotic membrane stem cells, one is enzymatic digestion, and the other is the method of crawling out tissue pieces. The tissue piece climbing out method generally cuts the amniotic membrane tissue pieces into pieces, and uses the climbing out method to connect them to the bottom of the petri dish. In the process, because the tissue block is too tightly attached to the culture dish, when removing the tissue, the removal is not clean, and the tissue block and cells will be mixed together, and an additional cell sieve filtration process is required to collect the cells; in addition, because the amnion epidermis is flaky , is relatively thin and sticky, and will stick together. It cannot be spread out when it is inserted into the culture dish, or it takes a long time to spread it out. The tissue blocks are very easy to overlap, which will affect the cells crawling out.

[0010] Therefore, there is a need for a new method for amnion stem cell extraction, which avoids overlapping of tissue pieces or mixing of tissue pieces with crawling cells, and it is not easy to collect cells

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