核移植介導(dǎo)生產(chǎn)轉(zhuǎn)基因延邊奶山羊供核細(xì)胞體系的研究
發(fā)布時(shí)間:2024-03-27 04:29
【文章頁數(shù)】:116 頁
【學(xué)位級別】:博士
【文章目錄】:
CHPATER 1 Establishment and characterization of the mammary gland epithelial cell line inYanbian dairy goat
Abstract
1 Introduction
2 Materials and Methods
2.1 Medium preparation for cell culture
2.2 Isolation and culture of DGMECs
2.3 Biological characteristics of DGMECs
2.4 Statistic analysis of data
3 Results
3.1 Morphology of DGMECs
3.2 Growth curve of DGMECs
3.3 Average adherence and survival rate of cell
3.4 Doubling time of cell population
3.5 The mitotic index of DGMECs
4 Discussion
References
CHAPTER 2 Establishment of an embryonic stem cell line from blastocyst embryos in mouse
Abstract
1 Introduction
2 Materials and Methods
2.1 Preparation of MEF
2.2 Mitomycin C-inactivation of MEF cells
2.3 Preparation of freeze stock
2.4 Mice, embryos and isolation of ES cells
2.5 Statistic analysis of data
3 Results and analysis
3.1 Characteristics of the MEF isolated from different embryonic mouse age
3.2 Influence of different trypsinase concentration on isolation of MEF
3.3 The growth curve of the mouse MEF
3.4 The mitotic index of the MEFs
3.5 Cryopreservation and recovery of MEF
3.6 The concentration and treatment time of MMC treatment on the MEFs
3.7 Effect of ES cell isolate by different treatment time with MMC
3.8 Effect of different MEF concentration on ICM and ES cells
3.9 Effecs of LIF concentration on the growth of mouse ES cell
4 Discussion
References
CHAPTER 3 Establishment of Embryonic Stem Cells From NT Blastocystin Yanbian Goat
ABSTRACT
1 INTRODUCTION
2 Materials and Methods
2.1 Preparation of MEF
2.2 Goat Fetal Fibroblast Feeder Cell Line
2.3 Embryo Collection
2.4 ES Cell Isolation
2.5 Differential Staining
2.6 Statistic analysis of data
3 RESULTS
3.1 Growth curve of goat fetal fibroblast Cell
3.2 Embryo Culture and ES Cells Derivation
4 DISCUSSION
REFERENCES
CHAPTER 4 Method of Establishing Embryonic Stem (ES) Cell line
1 Established ES cell culture system method
1.1 The feeder layer culture system
1.2 Feeder-free culture systems
2 Embryo collection
3 ICM's acquisition
3.1 Direct separation method
3.2 Immune separation method
3.3 Heat shock method
3.4 Delayed implantation method
3.5 ES cell tissue blocks culture
3.6 Feeder layer inverted culture
4 Endoderm group's Discretion
5 Identification of ES cells
REFERENCE
CHAPTER 5 The Normal Microenvironment In Mammary Gland Development
Abstract
Introduction
Role of Normal Mammary Microenvironment in Tissue Engineering
Conclusions and Future Perspectives
References
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