Unlock instant, AI-driven research and patent intelligence for your innovation.

Method for non-contact co-culture of mouse primary keratinocytes and primary immune cells

A keratinocyte, immune cell technology, applied in cell dissociation methods, epidermal cells/skin cells, cell culture supports/coatings, etc., can solve the physiological interference of cytokine release keratinocytes, terminal differentiation of keratinocytes, cell Problems such as low survival rate, to achieve the effect of convenient application of experimental factors, improvement of survival rate, and improvement of cell activity

Pending Publication Date: 2020-09-18
THE FIRST AFFILIATED HOSPITAL OF ANHUI MEDICAL UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing method for isolation of primary mouse keratinocytes has the problems of low cell survival rate, low cell purity, and other cell contamination
In addition, the complete use of serum-free medium suitable for keratinocytes in the co-culture system of primary keratinocytes and immune cells may cause the lack of components necessary for the growth of immune cells, affecting the growth and metabolism of immune cells, while completely using traditional media may cause Terminal differentiation of keratinocytes, which interferes with the release of related cytokines and keratinocyte physiology

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 non-contact co-culture of mouse primary keratinocytes and primary immune cells
  • Method for non-contact co-culture of mouse primary keratinocytes and primary immune cells
  • Method for non-contact co-culture of mouse primary keratinocytes and primary immune cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A method for non-contact co-cultivation of mouse primary keratinocytes and primary immune cells, comprising the following steps:

[0032] (1) Isolation of primary mouse keratinocytes: newborn mice placed in CO 2 The euthanasia chamber was executed for 20 minutes, and then immersed in 75% ethanol to remove microbial contamination on the skin surface; the limbs and tail were cut off, the skin of the trunk of the mouse was separated, cut into 4 small pieces, with the epidermis facing up and the dermis facing down, and immediately placed in HB SS ( Float the skin in Dispase II (2mg / mL) prepared without calcium and magnesium ions), and digest overnight at 4°C in the dark; use pointed tweezers to gently peel off the epidermis and put it in pre-cooled HBSS Wash to remove the remaining Dispase II on the epidermis; quickly put it into 5mL 0.05% trypsin (diluted in HBSS), digest it in a 37°C water bath for 5 minutes, shake it several times, then put it in a 37°C water bath for 5 mi...

Embodiment 2

[0046] A method for non-contact co-cultivation of mouse primary keratinocytes and primary immune cells, comprising the following steps:

[0047] (1) Isolation of primary mouse keratinocytes: newborn mice placed in CO 2 Killed in euthanasia chamber for 20 minutes, then soaked in 75% ethanol to remove microbial contamination on the skin surface; cut off the limbs and tail, separated the mouse trunk skin, cut it into 4 small pieces, with the epidermis facing up and the dermis facing down, and immediately put it into HB SS Float the skin in Dispase II (2mg / mL) prepared (without calcium and magnesium ions), and digest overnight at 4°C in the dark; use pointed tweezers to gently peel off the epidermis and put it in pre-cooled HBSS Wash in medium to remove the remaining Dispase II on the epidermis; quickly put it into 5mL 0.05% trypsin (diluted in HBSS), digest it in a water bath at 37°C for 5 minutes, shake it several times, then put it in a water bath at 37°C for 5 minutes; add 10...

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 non-contact co-culture of mouse primary keratinocytes and primary immune cells and belongs to the technical field of biology. The method comprises the steps of mouse primary keratinocyte separation, mouse primary keratinocyte culture, mouse skin immune cell separation, and non-contact co-culture of mouse primary keratinocytes and immune cells. According to themethod, a primary keratinocyte separation method is modified, and a perfect primary keratinocyte and primary immune cell co-culture system is established, so that the growth environment of the cells is simplified, experimental factors are convenient to apply, the experimental result is convenient to observe, and a technical approach is provided for study on changes of keratinocytes in immunodeficient mice.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for non-contact co-cultivation of mouse primary keratinocytes and primary immune cells. Background technique [0002] The method of preparing and culturing primary keratinocytes from neonatal and adult mouse epidermis is often used in the study of signaling pathways, differentiation and cell transformation in vitro, and the co-culture of primary mouse keratinocytes and primary immune cells is a good way to evaluate immunodeficiency. It is an important method for changing keratinocyte phenotype in mouse cell transplantation, and is of great significance for establishing mouse models of skin diseases. At present, the common method for isolating primary mouse keratinocytes is trypsin cold-warm two-step digestion method. Existing mouse primary keratinocyte isolation methods have the problems of low cell viability, low cell purity, and other cell contamination. In addition, the...

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): C12N5/071C12N5/078
CPCC12N5/0629C12N5/0634C12N2509/00C12N2500/90C12N2500/14C12N2533/54C12N2502/094C12N2502/11
Inventor 孙良丹俞亚芬甄琪李报雍亮葛荟瑶毛艺文陈微微
Owner THE FIRST AFFILIATED HOSPITAL OF ANHUI MEDICAL UNIV