Separation, culture and purification methods of human amniotic mesenchymal stem cells

Abstract

The invention provides a separation method of human amniotic mesenchymal stem cells. The separation method comprises the following steps: (1) collecting human placental tissues and separating amniotictissues; washing blood stains; (2) chopping the amniotic tissues and adding a first tissue digestion solution and carrying out digestion; (3) after finishing the digestion, removing the first tissuedigestion solution; washing the amniotic tissues; adding a second tissue digestion solution and carrying out digestion; (4) after finishing the digestion, adding a trypsin solution and carrying out digestion; (5) adding a stop solution and stopping the digestion; filtering to remove tissue blocks which are completely digested, collecting a cell suspension solution and centrifuging; (6) removing supernatant, adding a DMEM (Dulbecco's Modified Eagle Medium) and washing sediment; centrifuging again to obtain the separated human amniotic mesenchymal stem cells. The method provided by the inventionis simple and effective, can obtain high-purity human amniotic mesenchymal stem cells and has no influences on cell activity.

Description

technical field [0001] The invention belongs to the field of stem cells and regenerative medicine, in particular to a method for separating, cultivating and purifying human amniotic mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of pluripotent stem cells derived from mesoderm with high self-renewal and multi-directional differentiation capabilities. They can be cultured and expanded in vitro and have multi-directional differentiation capabilities under specific conditions, and can differentiate into fat. Cells, chondrocytes, bone cells, liver cells, and even laterally differentiate into nerve cells, epidermal cells, etc. MSCs have broad clinical application prospects and are the most important seed cells in the fields of regenerative medicine and tissue engineering. [0003] Mesenchymal stem cells were first discovered in bone marrow tissue. With the deepening of research, they have also been found in various tissues of the hum...

AI Technical Summary

Problems solved by technology

Most of the existing amnion epithelial cells are separated by trypsin digestion, which can remove most of the amnion epithelial cells, but the removal of epithelial cells is not complete, and they will still affect each other during normal culture, and trypsin is a broad-spectrum protein Digestive enzymes, the digestion process will cause great damage to the cells. It is difficult to accurately grasp the concentration of trypsin, digestion conditions and digestion time. If the enzyme concentration is too high or the digestion time is slightly longer, the tissue and cells may be seriously damaged, resulting in the activity of the isolated amniotic mesenchymal stem cells. Damaged, affecting later use

Images