Method for preparing human umbilical cord mesenchymal stem cell exosomes

A technology for stem cells and exosomes, applied in the field of preparation of human umbilical cord mesenchymal stem cell exosomes, can solve the problems of low efficiency of immunomagnetic beads method and chromatography method, unfavorable large-scale production, complicated operation, etc. The bacteria are free of mycoplasma contamination, easy to mass-produce, and the effect of high product safety

Pending Publication Date: 2019-06-14
JINAN PANSHENG BIOTECH
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Problems solved by technology

The sample obtained by density gradient centrifugation is relatively pure, but it is time-consuming and complicated to operate, which is not conducive to large-scale production; the immunomagnetic bead method a

Method used

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  • Method for preparing human umbilical cord mesenchymal stem cell exosomes
  • Method for preparing human umbilical cord mesenchymal stem cell exosomes
  • Method for preparing human umbilical cord mesenchymal stem cell exosomes

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Embodiment Construction

[0037] The present invention provides a method for preparing exosomes derived from umbilical cord mesenchymal stem cells, comprising the following steps:

[0038] 1. Culture and identification of umbilical cord mesenchymal stem cells

[0039] 1.1 Isolation and culture: Collect the umbilical cord of full-term cesarean section neonates under sterile conditions and process it within 4 hours. Cut the umbilical cord with scissors into small sections of about 2-3 cm / , and wash the umbilical cord repeatedly with saline containing penicillin-streptomycin to remove surface blood stains. Remove the three blood vessels and amniotic membrane, peel off the Huatong glue, and cut the Huatong glue to about 1mm with scissors 2 small pieces, spread evenly on T75cm 2 In a culture bottle (when laying the bottle, try to distribute the tissue pieces evenly, leave a gap in the middle, do not cut out air bubbles, which is not conducive to cell climbing out), place at 37°C, 5% CO 2 Cultivate in an ...

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Abstract

The invention discloses a method for preparing human umbilical cord mesenchymal stem cell exosomes. The method uses a newborn umbilical cord as a source of umbilical cord mesenchymal stem cells, usesa medium prepared by a fetal calf serum or a serum substitute in which exosome carried by itself is removedfor culturing, collects a supernatant, and further separates and extracts the exosomesby differential centrifugation, the purity is high, and the source of the umbilical cord mesenchymal stem cell exosomes is ensured; the exosomes is prepared by the differential centrifugation, the operationis simple, and the method is suitable for large-scale production; during the whole process, sterility is guaranteed, the contamination ofmycoplasma is prevented, and the product safety is higher.

Description

technical field [0001] The invention relates to a method for preparing human umbilical cord mesenchymal stem cell exosomes, belonging to the field of cell culture. Background technique [0002] Umbilical cord mesenchymal stem cells are mesenchymal stem cells derived from umbilical cord Huatong jelly tissue. It has a wide range of sources, low immunogenicity, and has multiple functions such as differentiation, immune regulation, and hematopoietic support. It has become the seed cell of cell therapy. It has good clinical efficacy in many diseases including diabetes, nervous system disease, liver disease, cardiovascular disease, autoimmune disease and other diseases, but its mechanism of action is still unclear. It was initially believed that umbilical cord mesenchymal stem cells would homing to the damaged site after transplantation into the human body for cell self-renewal and directional differentiation. The limited number of target cells does not adequately account for the...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 陈莉邢志青王红丽田晓婷朱继玲周健关美月
Owner JINAN PANSHENG BIOTECH
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