Method for construction of stem cell in-vitro multiplication and differentiation microenvironment

An in vitro proliferation and stem cell technology, applied in artificial cell constructs, non-embryonic pluripotent stem cells, biochemical equipment and methods, etc. The effect of promoting significant proliferation and delaying differentiation

Inactive Publication Date: 2007-05-16
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
View PDF0 Cites 23 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned approaches have the following problems: (1) The commodity price of cytokine LIF is expensive, which not only increases the harshness of ES cell maintenance culture conditions, but also greatly limits the extensive development of large-scale expansion of ES cells
(2) The preparation of feeder cells has high requirements on the source of materials (pregnant mice at 13.5 days pregnant), the operation is complicated, and feeder cells pretreated with mitomycin are required for each passage
(3) The transgenic operation of eukaryotic cells is relatively complicated, and the expression of foreign genes is unstable and easy to be lost in the process of proliferation and passage
However, in recent years, most of the relevant reports have focused on the in vivo induced differentiation of adult stem cells, and there are few studies on ES cells.

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 construction of stem cell in-vitro multiplication and differentiation microenvironment
  • Method for construction of stem cell in-vitro multiplication and differentiation microenvironment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0030] Reagents: sodium alginate (Sigma), polylysine (Sigma), monoclonal antibody SSEA-1 (SantaCruz), alkaline phosphatase AP (Zhongshan), RI-PCR kit (Takara);

[0031] ES cell growth medium: H-DMEM medium (Gibico) + 15% fetal bovine serum + 1% non-essential amino acids + 0.1mmol / L β-mercaptoethanol (β-Me) + 10 3 U / ml LIF;

[0032] Instrument: CO 2 Cell incubator, phase contrast microscope, electrostatic droplet generator, microplate reader, UV spectrophotometer, laser confocal microscope, stirred bioreactor (B.Braun);

[0033] Cell model: mouse embryonic stem cell line ES-D 3 (Provided by Peking University Health Science Center);

[0034] specific method:

[0035] 1. ES-D 3 Cells are maintained in vitro in growth medium. After being digested with 0.25% trypsin and collected, the cell density was adjusted to 4×10 6 cells / ml. Using the electrostatic droplet generation method, APA microencapsulated mouse ES cells were prepared under physiological conditions, and the spec...

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 a constructing method of stem cell breeding and differentiation microenvironment in vitro in the stem cell tissue engineering domain, which comprises the following steps: adopting sodium alginate and polylysine as material to prepare APA microencapsulation ES cell; adding microencapsulation ES cell in the ES growing culture medium; stewing at 37 Deg C in the culture box with 5% CO2; changing culture medium every 2-3d to obtain the product. The invention simplifies the operation to generate high-density and large scale augmentation, which can inhibit or delay the oriental differentiation for ES cell.

Description

technical field [0001] The invention relates to the field of stem cell tissue engineering, in particular to a method for constructing a microenvironment for proliferation and differentiation of stem cells in vitro. Background technique [0002] Embryonic stem cells (ESC) is a specific cell group derived from the inner cell mass of the blastocyst, which has the potential of self-renewal and omnidirectional differentiation, that is, under suitable in vivo or in vitro conditions, it can differentiate into Cells of three different germ layers: inner, middle, and outer. In 1981, the successful isolation and establishment of mouse ES cells opened the prelude to stem cell research [Document 1: Evans MJ, Kaufman MH. Establishment in culture of pluripotential cells from mouseembryos. Nature, 1981; 292(5819): 154-156 .]. Subsequently, the directional induction of ESC differentiation has become a hot spot of common concern at home and abroad. At present, the research on the directio...

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): C12N5/06C12N5/08C12N5/071C12N5/0735C12N5/074C12N5/0775C12N5/0789C12N5/0797
Inventor 马小军王秀丽王为
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap