Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Autologous hair growth matrix stem cell culture method for cerebral hemorrhage sequelae of premature infant

A technology for cerebral hemorrhage sequelae and stem cell culture, applied in the field of neural stem cell culture, can solve problems such as restriction and immune system rejection, and achieve the effect of promoting maturation, reducing neurological sequelae, and avoiding re-bleeding

Pending Publication Date: 2022-06-21
浙江以和细胞生物科技有限公司
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The use of neural stem cells to treat sequelae of cerebral hemorrhage is a hot research field today. In recent years, animal and clinical experiments of neural stem cell transplantation at home and abroad have confirmed that it has a certain effect on brain injury, providing a scientific basis for neural stem cell transplantation in the treatment of sequelae of cerebral hemorrhage. The lack of sources of neural stem cells and the rejection of the immune system after transplantation limit further research on the treatment of brain injury

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
  • Autologous hair growth matrix stem cell culture method for cerebral hemorrhage sequelae of premature infant
  • Autologous hair growth matrix stem cell culture method for cerebral hemorrhage sequelae of premature infant
  • Autologous hair growth matrix stem cell culture method for cerebral hemorrhage sequelae of premature infant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Irrigation of hematoma and collection of bloody cerebrospinal fluid under neuroendoscopy

[0059] Use ultrasound 3D navigation to accurately locate the lateral ventricle, design a small incision, cut the skin, cut the skull with scissors, and puncture the cortex under ultrasound guidance. The irrigation system has two irrigation channels, a scope channel, and a working channel, allowing simultaneous irrigation and collection of fluid.

[0060] After incision of the cortex, one channel is connected to the infusion tube and the other is connected to the opening of the third ventricle. The lavage speed is gravity-related (1.8m suspension), so there is no risk of accidental injury, including complications such as elevated blood pressure. In addition, neurosurgeons constantly monitor fontanelle tension and can open or close the circuit to maintain a steady pressure. The method of collecting cerebrospinal fluid is to use a 50 ml syringe connected to the outflow ch...

Embodiment 2

[0061] Example 2 Cell collection and culture

[0062] The CSF samples were transferred to appropriately sized tubes and centrifuged at 370g for 10 minutes, the supernatant was discarded, and the stem cells were resuspended in neural stem cell medium to maintain the culture at a seeding density of 1 × 10 4 -1×10 5 pcs / mL.

[0063] Neural stem cell culture medium consists of the following components: Neurobasal medium is a basal medium containing EGF 20ng / mL, bFGF 20ng / mL, LIF 10ng / mL, heparin 2μg / mL, B27 supplement (50×, Gibco) 2%, N2 additive (100×, Gibco) 1%, non-essential amino acids (glycine, L-alanine, L-asparagine, L-aspartic acid, glutamic acid, L-proline, L-serine) 0.1 mM each, poly-L-ornithine 20 μg / mL and human placental laminin 20 μg / mL.

[0064] The entire process was carried out in 3D very low adsorption culture flasks (Corning Incorporated, Corning, NY). The medium is changed in half every 3-5 days, and the passage is once every 6-8 days. The specific passage ...

Embodiment 3

[0065] Example 3 Cryopreservation and recovery of subcultured germinal stromal stem cells

[0066] Cryopreservation: Transfer the germinal stromal stem cells cultured to the fifth passage into a centrifuge tube, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, and use the Accutase enzyme solution with a volume ratio of 1:1 and 10 units / minute for the pellet. Digest the mixed digestion solution of the papain solution with mL of papain solution. The digestion time is about 5 minutes. Gently pipetting and then centrifuging at a centrifugal speed of 1000 rpm for 10 minutes. Overhang adjusted to 5×10 6 1 / mL, aliquot into cryopreservation tubes, seal the tubes tightly, place them in the freezer at 4°C for 20 minutes, then place them in the freezer at -20°C for 30 minutes, and then place them in a -30°C low-temperature freezer 1 hour, and finally placed in liquid nitrogen for cryopreservation. The cryopreservation solution is Neurobasal medium containing 8%-12% DMSO b...

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

No PUM Login to View More

Abstract

The invention discloses a method for culturing autologous hair-growing stromal stem cells for cerebral hemorrhage sequelae of a premature infant. The method comprises the following steps: 1) collecting bloody cerebrospinal fluid from a lateral ventricle of the cerebral hemorrhage premature infant; 2) centrifugally collecting cells; 3) resuspending the cells by using a neural stem cell culture medium and maintaining culture; 4) changing a half amount of the liquid in 3-5 days, digesting the cells after 6-8 days, and carrying out subculture; and 5) after passage to 3-5 generations, collecting the hair growth matrix stem cells. Through primary culture and subculture, the autologous neural stem cells of the child patient are massively amplified, then cryopreserved, recovered at a proper time and implanted into the brain of the child patient, so that the treatment effect on the cerebral hemorrhage sequelae of the premature infant can be greatly improved. The hair-growing stromal stem cells cultured by the culture method disclosed by the invention are large in amplification quantity and stable in property, and can meet the requirements of autotherapy of sick children.

Description

technical field [0001] The invention relates to a method for culturing neural stem cells, in particular to a method for culturing autologous germinal basement membrane stem cells for the sequelae of premature infants with cerebral hemorrhage. Background technique [0002] Neonatal intracerebral hemorrhage is different from adult intracerebral hemorrhage. All neonatal intracerebral hemorrhages are caused by the germinal stroma. The neonatal brain germinal matrix (GM) is rich in new blood vessels and newly migrated glial cells, while the GM blood vessels in preterm infants lack elastin and muscularis mucosae, and are prone to rupture and hemorrhage under vascular pressure changes. Germinal Matrix Hemorrhage (GMH) is one of the most serious neurological diseases in neonates, with an incidence of about 3.5‰. Children with GMH have many sequelae, including hemiplegia, aphasia, coma, and developmental disorders. Most children will develop the well-known cerebral palsy disease. ...

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
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797C12N5/079C12N5/0793A01N1/02
CPCC12N5/0623C12N5/0619C12N5/0622A01N1/0221C12N2509/00C12N2501/11C12N2501/115C12N2501/235C12N2533/32C12N2533/52C12N2500/32C12N2501/91C12N2500/38C12N2501/13C12N2501/15
Inventor 陈金虎
Owner 浙江以和细胞生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com