Mouse Embryonic Stem Cells Were Isolated And Cultured And Biological Characteristics Of Study

ObjectiveEmbryonic stem cells (ESCs) are pluripotent cells derived from the inner cell mass (ICM) of pre-implantation mammal or human embryos and primordial germ cells (PGCs) of embryos. ESCs were established as a suitable model to study the events which occurred in early development of mammalian embryo, such as cell differentiation and gene expression regulation. It is also an ideal source of tissue engineering and cell substitutive therapy of regenerative medicine. The target of this experiment is to investigate how to isolate totipotent mouse ESCs from BALB/C species mice and to analysis some biological characteristics of ESCs. Methods1, The blastocysts of 3.5d from BALB/C mice were collected, ICM were isolated and cultured on the primary mouse embryonic fibroblast (PMEF) cell feeder layers. The changes of cell colonies were observed by inverted phase contrast microscope and recorded.2,ES-like colonies were continued disaggregated and cultured. The ES-like colonies were identified by detection of some special markers of ESCs, alkaline phosphatase (AKP) and stage-specific embryonic antigen (SSEA) -1, and by observation the ability of differentiation in Vitro.3, Cytoplasmic fraction and nuclear fraction of ESCs in different stage were extracted and the relationship of the proliferation and differentiation of ESCs with the activation of nuclear factor kB (NF-κB) were analyzed. Results1,In this study, 71 blastcysts of BALB/C mice were collected, 30 ES-like colonies were obtained and most of them could be cultured to more than three passages, one of them was passaged to 12 passages and cultured stablely .2,All the ES-like colonies showed typical morphological characteristics of ESCs, AKP and SSEA-1 positive expression, and could differentiate to neuron-like...