The invention belongs to the technical field of
biological product separation culture, and particularly relates to a human
umbilical cord mesenchymal stem cell separation culture method. The method comprises the following steps that an in-vitro
umbilical cord of a term
fetus born by
caesarean delivery is taken and put into a sterile tissue preserving fluid for storage; flushing is conducted multiple times with normal
saline, and remained bloodstains are removed; two umbilical veins and two umbilical arteries of tissue
mass are removed with toothed
tweezers, and Wharton's jelly is completely
cut into pieces with sterile scissors; the Wharton's jelly which is
cut into pieces is transferred into a culture medium, and vibration and
centrifugation are conducted in sequence; the centrifugal
sedimentation part is transferred into a
cell culture flask; after culture is conducted for 5-6 days, it can be seen that part of cells
climb out from the periphery of small pieces of the tissue, then a culture substrate is replaced once
every three days, and culture continuous to be conducted; on the fourteenth day or so, the degree of
cell fusion reaches 80% or above, and spiral growth is achieved;afterwards, each transmission of the next generation takes three days, and adequate mesenchymal stem cells can be obtained. Compared with a traditional tissue method, the method is simpler, and the purity and yield of stem cells are higher.