Umbilical cord tissue mesenchymal stem cell isolated culture method

A technology of umbilical cord tissue and stem cells, which is applied in the field of stem cells and bioengineering, and can solve problems such as cell damage

Active Publication Date: 2016-01-13
SHENZHEN HORNETCORN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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Method used

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  • Umbilical cord tissue mesenchymal stem cell isolated culture method
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  • Umbilical cord tissue mesenchymal stem cell isolated culture method

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Sample Collection and Cell Isolation and Culture (The reagent consumables used in the experiment are all sterile)

[0034] 1. Collect the umbilical cord tissue of the postpartum full-term fetus under strict aseptic conditions, put it into a sterile glass bottle containing MSC medium supplemented with 100U / ml penicillin and 100μg / ml streptomycin, and transport it at 4°C. Make sure the umbilical cord tissue is completely submerged below the surface of the medium solution and process the umbilical cord tissue within 48 hours.

[0035] 2. Put the fetal umbilical cord tissue into a 15cm petri dish in an ultra-clean workbench, and use forceps to squeeze out the blood in the umbilical cord. The umbilical cord tissue was then washed with PBS until the blood on the surface of the umbilical cord was cleaned.

[0036] 3. Use scissors to cut the umbilical cord tissue into 3-5cm length, use forceps to tear the amniotic membrane on the surface of the umbilical cord, and remove the...

Embodiment 2

[0064] Sample collection and cell isolation and culture (the reagent consumables used in the experiment are all sterile)

[0065] 1. Collect the umbilical cord tissue of the postpartum full-term fetus under strict aseptic conditions, put it into a sterile glass bottle containing MSC medium supplemented with 100U / ml penicillin and 100μg / ml streptomycin, and transport it at 4°C. Make sure the umbilical cord tissue is completely submerged below the surface of the medium solution and process the umbilical cord tissue within 48 hours.

[0066] 2. Put the fetal umbilical cord tissue into a 15cm petri dish in an ultra-clean workbench, and use forceps to squeeze out the blood in the umbilical cord. The umbilical cord tissue was then washed with PBS until the blood on the surface of the umbilical cord was cleaned.

[0067] 3. Use scissors to cut the umbilical cord tissue into 3-5cm length, use forceps to tear the amniotic membrane on the surface of the umbilical cord, and remove the a...

Embodiment 3

[0075]Sample collection and cell isolation and culture (the reagent consumables used in the experiment are all sterile)

[0076] 1. Collect the umbilical cord tissue of the postpartum full-term fetus under strict aseptic conditions, put it into a sterile glass bottle containing MSC medium supplemented with 100U / ml penicillin and 100μg / ml streptomycin, and transport it at 4°C. Make sure the umbilical cord tissue is completely submerged below the surface of the medium solution and process the umbilical cord tissue within 48 hours.

[0077] 2. Put the fetal umbilical cord tissue into a 15cm petri dish in an ultra-clean workbench, and use forceps to squeeze out the blood in the umbilical cord. The umbilical cord tissue was then washed with PBS until the blood on the surface of the umbilical cord was cleaned.

[0078] 3. Use scissors to cut the umbilical cord tissue into 3-5cm length, use forceps to tear the amniotic membrane on the surface of the umbilical cord, and remove the ar...

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Abstract

The invention discloses an umbilical cord tissue mesenchymal stem cell isolated culture method. The method comprises the steps of A, sample collection and cell isolated culture; B, subculture, wherein after being cultured, primary cells are subcultured continuously when cell confluence reaches over 80%; C, growth curve drawing; D, cell detection, wherein the third generation of cells are adopted for detection, well digested third generation of umbilical cord mesenchymal stem cells are prepared and washed with a buffer solution, then antibody labeling is conducted on cells, incubation washing is conducted, and then detection is conducted. Two types of collagenase are used at the same time, and digestion of umbilical cord tissue is conducted with hyaluronidase and neutral protease or accutase, so that tissue blocks are quickly and fully digested.

Description

Technical field [0001] The invention involves the field of stem cells and biological engineering, which is specifically a separate culture method for mesenchymal stem cells between umbilical cords. technical background [0002] The germinated stem cells were separated from the bone marrow in the 1970s.Later, people found that this cell has multi -directional differentiation potential, and can be differentiated into osteoblasts, cartilage cells, fat cells, myocardial cells, etc. under certain induction and culture conditions.It can still maintain the potential of differentiation.Because it can differentiate into the megenic cells of multiple medium germ sources, it is called mesenchymalstemcell (MSC).With the deepening of research, people have discovered that accommodated stem cells not only have multiple differentiation potential, but also have certain disease treatment potential.At present, it has been found in many animal experiments that after transplanting mesenchymal stem ce...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 刘聪陈晨王宇环罗晓玲魏志璋
Owner SHENZHEN HORNETCORN BIOTECH
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