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

In-vitro three-dimensional culture method for palatal organ

A three-dimensional culture and organ technology, applied in the field of cell and molecular biology, to achieve the effect of simple operation and low cost

Active Publication Date: 2014-06-11
DALIAN MEDICAL UNIVERSITY
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no matter which culture method is used, the longest culture time of palatal plates in vitro is only 80 hours

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
  • In-vitro three-dimensional culture method for palatal organ
  • In-vitro three-dimensional culture method for palatal organ
  • In-vitro three-dimensional culture method for palatal organ

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Matrigel three-dimensional culture of palatal plate tissue in vitro

[0029] The middle one-third of the facial tissues (all the tissues from the infraorbital rim to the corner of the mouth, including the palate, the lower part of the nasal septum and surrounding tissues, referred to as palatal organs) were separated from the normal mouse embryonic palate development period (embryonic day 5). Mix with Matrigel Matrigel diluted 1-fold or 2-fold in serum-free DMEM, 10 samples per group, and place in an incubator to solidify into a gel. After 1 hour, DMEM medium containing 10% fetal bovine serum was added for incubation. Unembedded mouse palate tissue was used as the experimental control group (10 samples). All groups were cultured in vitro for 6 days, and fresh culture medium was replaced every two days. After 6 days, the palatal plate tissue of each group was observed, the number of samples with normal palatal plate tissue structure, bilateral palatal plate ...

Embodiment 2

[0035] Example 2: Collagen hydrogel three-dimensional culture of palatal plate tissue in vitro

[0036] Separation of tissues from the middle third of the palate during embryonic development of spontaneous cleft palate mice (day 6 of embryonic stage) (all tissues from the infraorbital rim to the corner of the mouth, including the palate, the lower part of the nasal septum and surrounding tissues, referred to as palatal organs) , and then mixed with mouse type II collagen (molecular weight 500kDa) solutions with a concentration of 5 mg / ml and 10 mg / ml respectively, 10 samples in each group, and put them in an incubator to solidify for 40 minutes to form a cylindrical hydrogel with a diameter of 2 cm and a thickness of 1 cm. Glue, and added DMEM medium containing 10% fetal bovine serum for culture. The unembedded palate tissue of mice with spontaneous cleft palate was used as the experimental control group (10 samples). All groups were cultured in vitro for 6 days, and fresh cu...

Embodiment 3

[0041] Example 3: In vitro three-dimensional culture of palatal plate tissue on oxidized hyaluronic acid / collagen composite hydrogel scaffold

[0042] Bovine sodium hyaluronate (molecular weight 1100 kDa) was prepared into a 10 mg / ml aqueous solution, and 20 mM sodium periodate solution was added according to the volume ratio of sodium hyaluronate / sodium periodate at 4:1, and stirred at 25°C in the dark. React for 5 hours, then dialyze, and finally freeze-dry to obtain oxidized hyaluronic acid.

[0043] Prepare 0.9% (w / v) collagen solution, and mix the collagen solution, DMEM medium and NaOH-NaHCO 3 -Hepes buffer is mixed according to the ratio of 7:2:1 (v / v), and then 0.6% (w / v) oxidized hyaluronic acid aqueous solution is mixed according to 10:1 and 30:1 (v / v) (solution: oxidized transparent Aqueous acid solution) was mixed, stirred slowly, and the mixture was placed in a refrigerator at 4 °C for 24 h.

[0044] Separation of the mid-third of the facial tissues of the mouse...

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

PropertyMeasurementUnit
Diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses an in-vitro three-dimensional culture method for a palatal organ. The method comprises the steps of taking all tissues from the infraorbital margin to the mouth angle part at the early development period or development period of an embryonic palate, and embedding in a hydrogel scaffold; adding a culture solution for three-dimensional culture. The method can provide an approximate in-vivo three-dimensional growth environment and avoid using expensive materials and processes, and has the advantages of simplicity in operation, low cost and the like. Moreover, the biological properties of palate plate tissues can be controlled by changing the composition of the hydrogel scaffold, and the method is suitable for culture of normal and abnormal palate plate tissues at different periods of embryonic development and can be used for research of a cleft palate formation mechanism and screening of palate plate forming promoting medicaments.

Description

technical field [0001] The invention relates to the fields of cell and molecular biology, in particular to a three-dimensional palatal organ culture method in vitro that can realize long-term high activity of palatal plate tissue. Background technique [0002] Cleft lip and palate are relatively common developmental deformities in newborns, and animals with cleft palate are difficult to survive after birth. In order to study the development process of the palate plate and the mechanism of the fusion of the palate plate, it is very necessary to culture the palate plate tissue in vitro. At present, there are only two methods of in vitro culture of palatal plate tissue: static culture and dynamic culture: the static culture method is to place the palate plate on a porous membrane, and add medium for static culture; the dynamic culture method is to suspend in the state of culture fluid flow to cultivate. However, no matter which culture method is used, the longest culture time ...

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
IPC IPC(8): C12N5/073
Inventor 肖晶孙广炜刘洋徐小溪李楠刘波丛蔚马小军
Owner DALIAN MEDICAL UNIVERSITY
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