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

Method for extracting unsaturated fatty acid composition from humen amniotic epithelial cells, and application thereof

A technology of epithelial cells and human amniotic membrane, applied in the separation/purification of carboxylic acid compounds, drug combinations, medical preparations containing active ingredients, etc., can solve the problem that the production of PUFAs cannot meet human needs, and there is no data showing human amniotic membrane epithelial cells Effectiveness, high price and other problems, to achieve the effect of reasonable proportion, broad medicinal prospect and low cost

Active Publication Date: 2014-04-16
广西南宁灵康赛诺科生物科技有限公司
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no data at home and abroad to show the role of human amniotic epithelial cells in this process
[0017] At present, PUFAs sold in the form of commercial products are mainly extracted from deep-sea fish, which are expensive and have limited yields. Microalgae and yeast strains that produce PUFAs through microbial genetic engineering also face many problems that need to be solved, making the production of PUFAs difficult. It has been unable to meet the needs of human beings, and it is urgent to develop new resources

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
  • Method for extracting unsaturated fatty acid composition from humen amniotic epithelial cells, and application thereof
  • Method for extracting unsaturated fatty acid composition from humen amniotic epithelial cells, and application thereof
  • Method for extracting unsaturated fatty acid composition from humen amniotic epithelial cells, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Morphological Observation and Lipid Staining of Amniotic Epithelial Cells of Normal Full-term Cesarean Section

[0037] The amniotic membrane was taken from a normal full-term cesarean section, fixed in 10% formaldehyde solution for 24 hours, rinsed in running water, dehydrated in gradient alcohol, transparent in xylene, embedded in wax. Slice with a LEICA microtome with a thickness of 7 μm. After spreading in warm water at 45°C, pick up the slides with poly-lysine-coated glass slides. Let dry and set aside.

[0038] Paraffin sections were routinely dewaxed to water. After washing with distilled water, dip in hematoxylin and lightly dye for 1-2 minutes, then wash with tap water. Add 70% alcohol for 5 seconds. Into Sudan III staining solution for 30 minutes. Wash with 70% alcohol for 5-10 seconds. Wash with distilled water, and cover with glycerin gelatin.

[0039] After the sections were stained in Nile blue sulfate solution for 30 minutes, they were s...

Embodiment 2

[0043] Example 2: Acquisition, isolation, cultivation and identification of human amniotic epithelial cells

[0044] The amniotic membrane was torn off from the placenta discarded after cesarean section, and placed in 1×PBS containing penicillin and streptomycin double antibodies and washed repeatedly until there was no blood. Spread the epithelium face down in a culture dish, and use a cell scraper to remove the connective tissue under the amnion dense layer. After repeated washing in 1×PBS, the amniotic membrane was cut into small pieces of about 1 mm×1 mm with ophthalmic scissors, added with newly prepared 0.01% collagenase IV, and placed in a 37°C incubator for 2-3 hours to digest. Collect the liquid in a centrifuge tube, centrifuge at 1000 rpm for 5 minutes, discard the supernatant. Rinse once with 1×PBS. Add 0.25% trypsin to digest at room temperature for 0.5-1 hour. Serum-containing medium terminates the digestion. Centrifuge at 1000 rpm for 5 minutes and discard th...

Embodiment 3

[0048] Example 3: Detection of the type and content of unsaturated fatty acids in free state extracted from human amniotic epithelial cells by gas chromatography / mass spectrometry

[0049] In order to reduce the interference of non-free fatty acids during the experiment, the cells were broken using a low-temperature ultra-high pressure continuous flow cell breaker, and 1×PBS 25ml was added to each 1.8L cell sample for dilution and washing. The whole process of breaking was in a low-temperature water bath at 4°C-6°C To maintain the activity and performance of the original substances in the cells. Add 75 ml of n-hexane to the crushed sample, and vibrate with a shaker at 500 rpm for 12 hours. After 25 ml of the organic phase was dried by nitrogen blowing, the dried fatty acid was derivatized with boron trifluoride methanol solution. After adding 10ml of n-heptane for extraction, take 5ml of it and dry it with nitrogen. Use 1ml of n-hexane to reconstitute and measure on the mach...

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 relates to a method for extracting an unsaturated fatty acid composition from humen amniotic epithelial cells (HAECs), and an application thereof. The unsaturated fatty acid composition mainly comprises polyunsaturated fatty acids (PUFAs) which comprise DHA, arachidonic acid, dihomo-gamma-linolenic acid, gama-linolenic acid and linoleic acid. The unsaturated fatty acid composition can be provided for preparing drugs for treating nervous system degenerative diseases (such as Parkinson's disease), ocular surface diseases, cardiovascular and cerebrovascular diseases and neural injury diseases. In the prior art, the commercial PUFAs are mainly extracted from deep-sea fishes or algaes, price of the PUFAs is expensive and output of the PUFAs is limit. According to the present invention, the free-state PUFAs composition extracted from the HAECs has advantages of sufficient source, low cost, high content, reasonable distribution ratio of n-6 PUFAs / n-3 PUFAs, and has a wide medicinal prospect.

Description

technical field [0001] The present invention relates to a method for isolating useful substances from human amniotic epithelial cells, in particular to a method and application for extracting unsaturated fatty acid compositions in a free state from the cytoplasm of human amniotic epithelial cells. Background technique [0002] Human amniotic epithelial cells are derived from placental amniotic membrane tissue. They are a kind of adult stem cells with obvious plasticity and multipotential differentiation potential. Under the regulation of different growth factors, they can differentiate into different tissue cell types derived from the three germ layers, such as liver Like cells, cardiomyocytes, glial cells, neurons and islet-like cells. During the early development of human embryos, human amniotic epithelial cells (HAECs) originate from the inner cell mass of the blastocyst, and are formed from some epiblast cells (amnioblasts) at the beginning of the second week after ferti...

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 Patents(China)
IPC IPC(8): C07C57/03C07C57/12C07C51/48A61K31/202A61K31/201A61P25/28A61P9/00
Inventor 朱梅陈彪
Owner 广西南宁灵康赛诺科生物科技有限公司
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