Method for separating and extracting hUC-MSC from umbilical cord outer layer amnion tissue

A technology for outer amniotic membrane and umbilical cord, applied to bone/connective tissue cells, vertebrate cells, animal cells, etc., which can solve the confusion of separation and amplification methods, complicated steps, and the difficulty of quickly extracting large quantities of high-purity hUC-MSCs And other issues
CN105420185AInactive Publication Date: 2016-03-23郭镭 +1
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
郭镭
Publication Date
2016-03-23
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a method for fast separating and extracting human Umbilical Cord mesenchymal stem cells (hUC-MSC) from umbilical cord outer layer amnion tissue. The method includes the following steps that fresh collected umbilical cord tissue of a healthy newborn is taken, transported on ice in double-antibody-containing umbilical cord storing transporting liquid, washed and disinfected with 75% ethyl alcohol and normal saline, outer layer amnions are separated bluntly after blood vessels are rejected, mechanically smashed and treated with erythrocyte lysate for 3 min, IV type collagenase digestion is used, and a digestion product is cultured in a suspension mode with a serum-free culture medium after being screened through a 100-200-mesh sieve; medium change is carried out every three to five days, supernatant is taken to detect cell contamination, after the attachment rate in a plate reaches 30-70% and trypsinization is used, cell passage amplification is collected centrifugally, and confluence is carried out when the cell confluence rate reaches 90% or above, the cells are collected for cryopreservation, and the biological characteristics of hUC-MSC are detected.
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Description

technical field

[0001] The invention relates to an efficient and rapid separation and extraction method for umbilical cord mesenchymal stem cells, in particular to a method for extracting mesenchymal stem cells from fresh umbilical cord outer amnion tissue. Background technique

[0002] Mesenchymal stem cells (mesenchymal stem cells, MSCs) are derived from the mesoderm and ectoderm in the early stages of development, have high self-renewal and multi-lineage differentiation potentials, and are widely distributed in connective tissues and interstitium of organs throughout the body. At present, studies have shown that different culture methods can cause MSCs to exhibit neuronal phenotypes, islet-like cell phenotypes, endothelial cell phenotypes, or express cardiomyocyte markers, thereby revealing the role of MSCs in immunosuppression, endocrine system regulation, and nervous system. It has broad clinical application prospects in regulating and improving cardiovascular function....

Claims

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