Application of human umbilical cord mesenchymal stem cells in preparing drugs for treating cerebral palsy and culture method

A technology of stem cells and culture method, which is applied to the application and culture of human umbilical cord mesenchymal stem cells in the preparation of cerebral palsy drugs, can solve problems such as little effect, and achieve the effects of less trauma, good patient compliance, and large cell volume

Inactive Publication Date: 2019-03-19
SHIYAN TAIHE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Traditional treatment methods are mainly supportive care. Traditional cerebral palsy treatment methods are usually used clinically, such as rehabilitation training, hyperbaric oxygen, neurotrophic drugs, cytokines, acupuncture, etc. These treatment methods are effective for nerve cells with relatively mild damage It has a certain effect, but for children older than 3 years old and / or for severely damaged nerve cells, it has little effect. The key is that all the treatment methods do not target the nerve repair of the brain lesion itself and / or intervention programs

Method used

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  • Application of human umbilical cord mesenchymal stem cells in preparing drugs for treating cerebral palsy and culture method
  • Application of human umbilical cord mesenchymal stem cells in preparing drugs for treating cerebral palsy and culture method
  • Application of human umbilical cord mesenchymal stem cells in preparing drugs for treating cerebral palsy and culture method

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Effect test

Embodiment 1

[0038] This embodiment provides the application and culture method of human umbilical cord mesenchymal stem cells in the preparation of cerebral palsy medicine. The specific treatment method includes the following steps:

[0039] (1) Take the human umbilical cord ligated after delivery of the fetus of a puerpera without infectious diseases, wash it with a sterilized PBS solution, place it in a PBS solution containing 900 U / mL gentamicin, and store it at a constant temperature at 2°C;

[0040] (2) the Wharton's colloid obtained after the separation of the human umbilical cord is cut into 1-4mm 3 The tissue homogenate block was added to the serum-free medium of human umbilical cord mesenchymal stem cells for culturing under the conditions of 3% carbon dioxide, constant temperature of 36°C, and humidity of 90%, to obtain cell clones;

[0041] (3) When the cell cloning group grows to 70% of the area of ​​the culture bottle wall, it is digested with 0.2% trypsin for 1 min, and the ...

Embodiment 2

[0048] This embodiment provides the application and culture method of human umbilical cord mesenchymal stem cells in the preparation of cerebral palsy medicine. The specific treatment method includes the following steps:

[0049] (1) Take the human umbilical cord ligated after delivery of the fetus of a puerpera without infectious diseases, wash it with a sterilized PBS solution, place it in a PBS solution containing 1100 U / mL gentamicin, and store it at a constant temperature at 8°C;

[0050] (2) the Wharton's colloid obtained after the separation of the human umbilical cord is cut into 4mm 3 The tissue homogenate was added to the serum-free medium of human umbilical cord mesenchymal stem cells for culture, the culture conditions were 7% carbon dioxide, 38°C constant temperature, and 100% humidity to obtain cell clones;

[0051] (3) When the cell cloning group grows to an area accounting for 90% of the wall area of ​​the culture bottle, it is digested with 0.3% trypsin for 3 ...

Embodiment 3

[0058] This embodiment provides the application and culture method of human umbilical cord mesenchymal stem cells in the preparation of cerebral palsy medicine. The specific treatment method includes the following steps:

[0059] (1) Take the human umbilical cord ligated after delivery of the fetus of a puerpera without infectious diseases, wash it with a sterilized PBS solution, place it in a PBS solution containing 1000 U / mL gentamicin, and store it at a constant temperature at 6°C;

[0060] (2) the Wharton's colloid obtained after the separation of the human umbilical cord is cut into 2mm 3 The tissue homogenate was added to the serum-free medium of human umbilical cord mesenchymal stem cells for culture, the culture conditions were 5% carbon dioxide, 37°C constant temperature, and 95% humidity to obtain cell clones;

[0061] (3) When the cell cloning group grows to 80% of the area of ​​the culture bottle wall, it is digested with 0.25% trypsin for 2 minutes, and the volume...

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Abstract

The invention relates to an application of human umbilical cord mesenchymal stem cells in preparing drugs for treating cerebral palsy and a culture method. The culture method comprises the following steps: culturing a Wharton jelly obtained from a human umbilical cord to obtain a cell clone group; then carrying out trypsin digestion on the obtained cell clone group to obtain P0 generation cells; carrying out subculture on the P0 generation cells to P4 generation; and identifying and screening surface antigens of P4 generation cells to obtain qualified human umbilical cord mesenchymal stem cells. The human umbilical cord mesenchymal stem cells are good in tissue integration, weak in immunogenicity and good in migrating ability, can be amplified quickly in vivo, has a multidirectional differentiative potential, can be differentiated to nerve cell directions, and can be applied to preparing drugs for treating cerebral palsy of children. The method is simple, safe, small in trauma and goodin compliance of patients, and preparation and clinical application standards and operating rules of human umbilical cord mesenchymal stem cells in preparing drugs for treating cerebral palsy of children are established, so that the culture method is safe and effective and has a good application prospect.

Description

technical field [0001] The invention belongs to the technical field of stem cell culture and application, in particular to the application and culture method of human umbilical cord mesenchymal stem cells in the preparation of cerebral palsy medicine. Background technique [0002] Cerebral palsy (CP) in children refers to central motor dysfunction and abnormal posture caused by non-progressive brain damage caused by various reasons from before birth to one year after birth, often accompanied by mental retardation and language barriers , epilepsy, low hearing, abnormal vision, etc., there is no effective clinical treatment. The etiology of this disease is relatively complex, and its main cause is premature birth, birth trauma, perinatal asphyxia and kernicterus, which cause cerebral ischemia and hypoxia in neonates. [0003] Traditional treatment methods are mainly supportive care. Traditional cerebral palsy treatment methods are usually used clinically, such as rehabilitati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K35/28A61P25/00
CPCA61K35/28A61P25/00C12N5/0665C12N2500/02C12N2509/00
Inventor 张澈顾教伟
Owner SHIYAN TAIHE HOSPITAL
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