Culture method of cd200+ subpopulation umbilical cord mesenchymal stem cells and medium used therefor

Abstract

The invention relates to the field of biomedicine, in particular to a method for culturing CD200+ subpopulation umbilical cord mesenchymal stem cells and a culture medium used therefor. The medium is a DMEM / F12 medium supplemented with the following components: 8v / v%~12v / v%FBS, FGF 8ng / ml~12ng / ml, BDNF 10ng / ml~30ng / ml and NT3 10ng / ml~30ng / ml ml. Using this medium for CD200 + The culture of subpopulations of umbilical cord mesenchymal stem cells can greatly accelerate the repair of spinal cord injury.

Description

technical field

[0001] The invention relates to the field of biomedicine, in particular to a method for culturing CD200+ subpopulation umbilical cord mesenchymal stem cells and a culture medium used therein. Background technique

[0002] The term "Spinal Cord Injury" refers to damage to the spinal cord caused by trauma (such as a car crash), disease, or degeneration (such as cancer). There are no reliable estimates of the global prevalence, but most are due to trauma, although the proportion of non-traumatic SCI is increasing. Symptoms of a spinal cord injury depend on the severity of the injury and the location of the damaged spinal cord. Symptoms may include partial or complete loss of sensory function or motor control of the arms, legs, and / or body. The most severe forms of spinal cord injury affect the body systems that regulate bladder and bowel control, breathing, heart rate, and blood pressure. Most people with spinal cord injuries experience chronic pain, and spin...

Examples

Embodiment 1

[0038] 1. CD200 + Preparation of Subgroup Umbilical Cord Mesenchymal Stem Cells

[0039] 1. Cultivation of Seed Cells

[0040] The sample cells used in this experiment were the 4th generation cells isolated from hUCMSC culture (Guangdong Province, China Umbilical Cord Blood Hematopoietic Stem Cell Bank). After removing the original medium, wash the cells with phosphate-buffered saline (PBS) for 1-2 times, and remove the remaining medium. Add an appropriate amount of DMEM / F12 medium for culture, observe the cell proliferation ability and morphological characteristics; proceed to the next step after several passages as required.

[0041] 2. Preparation of Cell Suspension

[0042] When the cells are cultured to close to 80% confluence, remove the medium, wash with PBS containing double antibodies for 1-3 times, add 0.125% trypsin, digest at 37°C for 10 minutes, add the same volume of medium to stop the digestion, repeat pipetting, and sieve through a 200-mesh sieve Filter with ...

technical field

[0001] The invention relates to the field of biomedicine, in particular to a method for culturing CD200+ subpopulation umbilical cord mesenchymal stem cells and a culture medium used therein. Background technique

[0002] The term "Spinal Cord Injury" refers to damage to the spinal cord caused by trauma (such as a car crash), disease, or degeneration (such as cancer). There are no reliable estimates of the global prevalence, but most are due to trauma, although the proportion of non-traumatic SCI is increasing. Symptoms of a spinal cord injury depend on the severity of the injury and the location of the damaged spinal cord. Symptoms may include partial or complete loss of sensory function or motor control of the arms, legs, and / or body. The most severe forms of spinal cord injury affect the body systems that regulate bladder and bowel control, breathing, heart rate, and blood pressure. Most people with spinal cord injuries experience chronic pain, and spin...

Examples

Embodiment 1

[0038] 1. CD200 + Preparation of Subgroup Umbilical Cord Mesenchymal Stem Cells

[0039] 1. Cultivation of Seed Cells

[0040] The sample cells used in this experiment were the 4th generation cells isolated from hUCMSC culture (Guangdong Province, China Umbilical Cord Blood Hematopoietic Stem Cell Bank). After removing the original medium, wash the cells with phosphate-buffered saline (PBS) for 1-2 times, and remove the remaining medium. Add an appropriate amount of DMEM / F12 medium for culture, observe the cell proliferation ability and morphological characteristics; proceed to the next step after several passages as required.

[0041] 2. Preparation of Cell Suspension

[0042] When the cells are cultured to close to 80% confluence, remove the medium, wash with PBS containing double antibodies for 1-3 times, add 0.125% trypsin, digest at 37°C for 10 minutes, add the same volume of medium to stop the digestion, repeat pipetting, and sieve through a 200-mesh sieve Filter with ...

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