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Method for separating and purifying totipotential stem cells of placenta

A totipotent stem cell, separation and purification technology, applied in cell dissociation methods, non-embryonic pluripotent stem cells, embryonic cells, etc., can solve the problems of cumbersome process and high separation cost, and achieve simple separation process, low separation cost and high separation efficiency. Effect

Inactive Publication Date: 2017-12-08
HENAN YINFENG BIOENG CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to provide a method for separating and purifying placental totipotent stem cells, so as to solve the existing problems of cumbersome process and high separation cost of placental totipotent stem cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A method for separating and purifying placental totipotent stem cells, comprising the steps of:

[0025] (1) Take the freshly collected human placenta tissue, wash it, put it into the placenta protection solution and save it for later use; wherein, the placenta protection solution is AIM-V serum-free medium;

[0026] (2) Cut the placental villi, and then cut the placenta into pieces; specifically: cut off the tissue with a thickness of 1 to 2 cm on the membrane surface of the placenta, and then cut it into pieces with a particle size not greater than 1 mm;

[0027] (3) Rinse with PBS buffer solution obtained in step (2) until colorless, digest in a water bath, shake and let stand for 5 minutes, so that the liquid is naturally divided into three layers;

[0028] (4) Draw the suspension of the intermediate layer into a centrifuge tube, add PBS buffer, shake well and centrifuge at 1000r / min for 5 minutes, discard the supernatant, and add the totipotent stem cell culture me...

Embodiment 2

[0031] A method for separating and purifying placental totipotent stem cells, comprising the steps of:

[0032] (1) Take the freshly collected human placenta tissue, wash it, put it into the placenta protection solution and save it for later use; wherein, the placenta protection solution is AIM-V serum-free medium;

[0033] (2) Cut the placental villi, and then cut the placenta into pieces; specifically: cut off the tissue with a thickness of 1 to 2 cm on the membrane surface of the placenta, and then cut it into pieces with a particle size not greater than 1 mm;

[0034] (3) Rinse with PBS buffer solution obtained in step (2) until colorless, digest in a water bath, shake and let stand for 15 minutes, so that the liquid is naturally divided into three layers;

[0035] (4) Draw the suspension of the intermediate layer into a centrifuge tube, add PBS buffer, shake well and centrifuge at a speed of 500r / min for 15 minutes, discard the supernatant, and add the totipotent stem cel...

Embodiment 3

[0038] A method for separating and purifying placental totipotent stem cells, comprising the steps of:

[0039] (1) Take the freshly collected human placenta tissue, wash it, put it into the placenta protection solution and save it for later use; wherein, the placenta protection solution is AIM-V serum-free medium;

[0040] (2) Cut the placental villi, and then cut the placenta into pieces; specifically: cut off the tissue with a thickness of 1 to 2 cm on the membrane surface of the placenta, and then cut it into pieces with a particle size not greater than 1 mm;

[0041] (3) Rinse with PBS buffer solution obtained in step (2) until colorless, digest in a water bath, shake and let stand for 10 minutes, so that the liquid is naturally divided into three layers;

[0042] (4) Draw the suspension of the intermediate layer into a centrifuge tube, add PBS buffer, shake well, and centrifuge at a speed of 500r / min for 10 minutes, discard the supernatant, and add the totipotent stem ce...

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Abstract

The invention discloses a method for separating and purifying totipotential stem cells of placenta. The method comprises the following steps: (1) washing a freshly collected human placenta tissue, putting into a placenta protecting solution, and storing for later use; (2) clipping placental villus, and cutting the placenta into pieces; (3) rinsing the PBS buffer solution obtained in the step (2) to be colorless, digesting in water bath, oscillating and standing to enable the liquid to be naturally separated into three layers; (4) pumping the intermediate suspension in a centrifugal pipe, adding the PBS buffer solution, uniformly shaking, centrifuging, removing supernatant, adding a totipotential stem cell culture solution to resuspend and precipitate; and (5) centrifuging, removing supernatant, adding the culture solution, uniformly mixing, transferring into a cell culture bottle, and culturing in a 5% CO2 culture tank at 37 DEG C. The method can be used for separating totipotential stem cells from placenta, is simple in separating process and convenient to operate, and the separated cell has high activity along with high separation cost and high separation efficiency.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for isolating placental totipotent stem cells. Background technique [0002] Placental totipotent stem cells are a type of totipotent stem cells obtained from placental tissue. Placental totipotent stem cells have the following characteristics: [0003] 1. It has strong proliferation ability and multi-directional differentiation potential, and can differentiate into mesenchymal stem cells, epithelial stem cells, neural stem cells, liver stem cells, muscle cells, osteoblasts, chondrocytes, and stromal cells under suitable in vivo or in vitro environments abilities of various cells. It can be used to repair damaged or diseased tissues and organs, and treat heart disease, cerebrovascular disease, nervous system disease, liver disease, bone tissue disease, corneal injury, burns, myopathy and other diseases. [0004] 2. It has the function of immune regulation. Through the nega...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074
CPCC12N5/0606C12N2509/00
Inventor 朱学义魏天迪王彦王秋文聂江华王豪东
Owner HENAN YINFENG BIOENG CO LTD
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