Studies On Human Amniotic Epithelial Cells Transfected With EGFP Transplantation Into The Lateral Ventricle Of Alzheimer's Disease Mice

Objective:(1) Propagation and identification of Alzheimer's disease model mice APPSWE/PS1DE9 transfer pairs of genes for further study to provide an ideal animal model.(2) To investigate the survival and differentiation of HAECs after being transplanted into the lateral ventricle of APPswe/ PS1DE9 transgenic mice model.(3) To investigate the effects of HAECs on behavior, pathology and cholinergic neurons acetylcholine neurotransmitters levels in the hippocampus of APPswe/PS1dE9 mice.Methods:(1) Parent generation and offspring mice were detected using PCR identification of APPSWE and PS1DE9 gene, Water maze and clear black-box were used to observation behavior by, Congo red and six ammonia silver staining were used to observation pathological changes .(2) With the EGFP labeled HAECs were transplanted into the lateral ventricle of mice, fluorescence microscopy was used to observation of HAECs survival and migration, immunohistochemistry was used to detect the expressin of Oct-4 and Nanog on HAECs.(3) Identification of the gene after the transgenic model and the negative models then were divided into four groups: sham-operated group model, the model treatment group, wild-type sham-operated group, wild-type treatment group.Treatment group, intracerebroventricular injection of HAECs, sham-operated group intracerebroventricular injection of the same amount of PBS. 5 weeks after transplantation, water maze and clear black-box observation of behavior change;HPLC electrochemical instrument observation of the content of acetylcholine in hippocampus; acetylcholinesterase staining of cholinergic neurons and nerve fibers of the change.Results:(1) Male and female mice APPswe/PS1 interactive, 32d within the middle eight nest 60, all survived. PCR identification of the mother on behalf of the 16 gene expression in mice APPswe positive; offspring 60 of 43 mice were positive, 17 gene expression is negative, the positive rate was 70.2%, survival rate was 100%. One APP gene fragment size 300bp. Transgenic mice compared with wild-type behavior are statistically significant difference (p<0.05). Transgenic rat model in August in the hippocampus and cortex occurs when amyloid deposition was dyed orange-red, wild-type mice did not find amyloid deposition.Two groups were not found in neurofibrillary tangles.(2) HAECs in the lateral ventricle in Alzheimer's disease model mice 4 weeks after transplantation could survive and migrate to the third ventricle, and express specific stem cell marker Oct-4 and Nanog.(3) 5weeks after transplantation, behavioral observation of transgenic mice treated group and the sham-operated group was statistically significant (p<0.01), normal wild-type aged rats sham-operated group and treatment group no significant difference (p<0.05). Transgenic mice and normal wild-type treatment group was no statistical difference in aged rats (p<0.05); transgenic model group significantly fewer neurons in the number of fibers with wild-type mice, transgenic model group, treatment group than in sham-operated group neurofibrillary increase in the number; transgenic mice treated group and the sham-operated group were statistically compared with acetylcholine content of different (p<0.05),Transgenic mice and wild-type mice treated group showed no significant difference (p<0.05). Wild-type mice treated group and the sham-operated group showed no significant difference (p<0.05). Conclusions:1. APPSWE/PS1DE9 transfer pairs of genes of Alzheimer's disease model mice can be stably inherited through the male and female mating continues, both in behavior and pathology with wild-type mice showed a significant difference.2. HAECs have stem cell properties, and can survive in the ventricles.3. Intracerebroventricular transplantation of human amniotic cells in Alzheimer's disease mouse model APPSWE/PS1DE9 transfer pairs of genes have therapeutic effects. To improve the symptoms of AD rats and its mechanism may be related to human amniotic epithelial cells release neurotrophic factor on neuronal protection, survival, more cholinergic neurons, the increase in cholinergic neurons in the hippocampus an increase of acetylcholine .