The Relationship Between Proliferation And Neural Differentiation Of Adipose Derived Mesenchymal Stem Cells And Expression Of P16ink4a Gene

Background:Alzheimer’s disease is one of the primary diseases threatening the life quality of aged people and its pathogenesis is complicated, so there is still no valid cure method. The autotransplantation of a rich source of easily accessible adipose derived mesenchymal stem cells, ADSCs, have enormous applying potential in the cure of Alzheimer’s disease. Whereas, the hypofunction of stem cells accompanied with Mammalian advancing age is a tough thing confronted by autologous stem cell transplantation. The genes that inhibit the cancer-p16ink4a is closely related with the senility of ADSCs and the excessive express of the genes exist in the Alzheimer’s disease, blocking-up of the genes of p16ink4a may help improve the inhibition of stem cell function caused by physiological senescence. To enhance the curative effect brought about by autotransplantation of ADSCs, it’s necessary to testify whether ADSCs have the age-related senescence and to observe the express change of the genes of pl6ink4a existing in the ADSCs of aged people.The experimental group infers that the ability of cell multiplication and directional differentiation of ADSCs weakens with the age, the expression of the genes of p16ink4a in cells increases and the genes of p16ink4a may affect cell multiplication and directional differentiation of ADSCs.Objective:To isolate ADSCs from young health rat adipose tissue as well as the old ones, inspect to compare the morphology, proliferation, neural differentiation of the two groups ADSCS and so on. Try to figure out the difference of expression of p16ink4a gene between both of the two groups. Methods:Inguinal and postperitoneal fat tissue was gotten from young/old rat, then to ADSCs were isolated and cultivated in media, and their shape was recorded. On confluence, the cells were passed. The third passage was put into gauge for growth curve by MTT, the cells were also identified by stem cells common marker CD29、CD90、CD11b with immunoflu-orescence stain. ADSCs were induced to neurocytes with the two stages strategy, and then cell expression of the neuron specific markers Nestin、Tubulin and GFAP was determined. SA β-gal staining were carried out for inspection the senescence of stem cells. Western Blot and real time PCR were implemented for the difference of p16ink4a expression in young ADSCs and Old rat derived ADSCs.Results:Indeed, ADSCs were reaped from rat fat tissue, and they showed a fusiform shape after stretching and rapid adhesion. It seemed that they could also pass infinitely in a considerable large amount. ADSCs strongly expressed CD29/CD90and did not express CD11b that can designate a stem cell. After a corporate induction with NPMM and BME, most of induced cells expressed Nestin which also figured the neuron-shape of these cells. Compared with the old group ADSCs, the young group ADSCs increased more rapidly, transformed to neurons more and expressed less p16ink4a gene.Conclusion:1. Young group ADSCs proliferated faster than the old ones and developed more neuron than the old one.2. Senile cells appeared more in the old rat adipose derived stem cells,while the expression of p16ink4a gene was also higher in the old group than the young ones.