Unlock instant, AI-driven research and patent intelligence for your innovation.
A method for isolating and culturing human umbilical cord mesenchymal stem cells
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A technique for stromal stem cells and a culture method, which is applied in the field of mesenchymal stem cell preparation, can solve the problem of the need to improve the purity of umbilical cord mesenchymal stem cells, and achieve the effects of fast expansion and cultivation, improved product quality and high purity.
Active Publication Date: 2020-10-27
浙江译美生物科技有限公司
View PDF1 Cites 0 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
[0004] In the study of cell phenotype identification, it was found that the purity of umbilical cord mesenchymal stem cells isolated and cultured by this method still needs to be improved
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more
Image
Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0038] Embodiment 1: the preparation of various raw materials
[0039] (1) Umbilical cord: collected from the obstetrics and gynecology delivery room of the hospital.
[0040] (2) PBS solution containing 100U / ml penicillin and 100U / ml streptomycin:
[0041] Weigh 1.42g Na 2 HPO 4 , 0.24g KH 2 PO 4 , 8g NaCl and 0.2g KCl, add 800ml deionized water and stir to dissolve, add 0.06g penicillin sodium and 0.1g streptomycin, adjust the pH value of the solution to 7.4 with HCl, and finally add deionized water to 1L, high temperature and high pressure Sterilize under the environment, obtain, store at room temperature; Penicillin and streptomycin in the solution are converted into valences of 100U / ml and 100U / ml respectively;
[0042] Among them, penicillin sodium and streptomycin were purchased from Gibco.
[0043] (3) Soaking solution:
[0044] Weigh a certain amount of glucose, (partially added with L-glutamic acid,) ethanol as required, add part of deionized water to stir and...
Embodiment 2-4
[0056] Embodiment 2-4: A method for isolating and culturing human umbilical cord mesenchymal stem cells, comprising the following steps:
[0057] i. Take the fresh isolated umbilical cord of the newborn, place it in PBS solution containing 100U / ml penicillin and 100U / ml streptomycin at a temperature of 2-5°C, and store it for later use;
[0058] ii. Take out the umbilical cord treated in step i, and wash it with PBS solution containing 100U / ml penicillin and 100U / ml streptomycin at a temperature of 2-5°C for several times until no blood remains;
[0059] iii. In the temperature-controlled soaking solution environment of 2-5°C, cut the washed umbilical cord, and remove the arteries and veins during the cutting process; the dosage ratio of the umbilical cord and the soaking solution is 1g:10-20ml, The solution includes 0.9-1.0g / L of glucose, 0.2-0.5g / L of L-glutamic acid, 10-20g / L of ethanol, and the balance is water;
[0060] iv. Take the cut umbilical cord, wash it 2-3 times ...
Embodiment 5
[0073] Example 5: A method for isolating and culturing human umbilical cord mesenchymal stem cells. The difference from Example 1 is that in Step IX and Step XI, the basal medium of the LG-DMEM medium does not contain HEPES.
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More
PUM
Login to View More
Abstract
The invention discloses a method for separating and culturing human umbilical cord mesenchymal stem cells. The method includes steps of acquiring fresh in-vitro umbilical cords of neonates; storing and washing the in-vitro umbilical cords by the aid of low-temperature PBS (phosphate buffer solution); shearing the in-vitro umbilical cords in low-temperature soak solution; washing the in-vitro umbilical cords by the aid of low-temperature PBS and carrying out low-temperature vacuum treatment on the in-vitro umbilical cords; thawing the in-vitro umbilical cords; adding the in-vitro umbilical cords into PBS and infiltrating the in-vitro umbilical cords; adding the in-vitro umbilical cords into first mixed enzyme solution and carrying out oscillation enzymatic hydrolysis treatment on the in-vitro umbilical cords; adding the in-vitro umbilical cords into second mixed enzyme solution and carrying out oscillation enzymatic hydrolysis treatment on the in-vitro umbilical cords; adding fetal calf serum, PBS and LG-DMEM (low glucose-dulbecco's modified eagle media) into the in-vitro umbilical cords to obtain first mixtures, allowing the first mixtures to stand still, then centrifuging the first mixtures and discarding supernatant; adding PBS into the first mixtures to obtain second mixtures, allowing the second mixtures to stand still, then centrifuging the second mixtures and discarding supernatant; adding fetal calf serum, PBS and LG-DMEM into the second mixtures to obtain third mixtures, allowing the third mixtures to stand still, then centrifuging the third mixtures and discarding supernatant; adding fetal calf serum, PBS and LG-DMEM into the third mixtures and uniformly mixing the fetal calf serum, the PBS, the LG-DMEM and the third mixtures with one another to obtain fourth mixtures; carrying out amplification culture on the fourth mixtures. The method has the advantage that the purity of the umbilical cord mesenchymal stem cells can be improved.
Description
technical field [0001] The invention relates to the preparation technology of mesenchymal stem cells, in particular to a method for separating and culturing human umbilical cord mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are a kind of pluripotent stem cells with high self-renewal ability and multi-lineage differentiation potential derived from the mesoderm and ectoderm in the early stage of development, which can be continuously subcultured under suitable conditions in vitro, Freeze. Because MSCs can differentiate into different cells under specific conditions, they have gradually become a very valuable cell source in the field of cell therapy and gene therapy. The umbilical cord belongs to the extra-embryonic tissue, which contains a large number of multidirectional stem cells, and the cell growth and expansion speed is fast, and the umbilical cord becomes "waste" after the fetus is delivered, so it is convenient to obtain materials...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More
Application Information
Patent Timeline
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to View More 
Login to View More