Study On Anti-aging Effect Of Mouse Fetus And Placenta Derived Cells Suspension And Mesenchymal Stem Cells

Objectives:To explore the anti-aging effect of the cells suspension and mesenchymal stem cells derived from murine fetus and placenta.Methods:1. Preparation of cells suspension of mouse fetus and placenta as well as isolation and identification of mesenchymal stem cells derived from them.We got 6-day-old Balb/c mouse fetuses and 18-day-old Balb/c mouse placentas (female and male) with uterine-incision delivery. The tissue cells suspension of mouse fetus and placenta were prepared with repeated blow and inhalation, grind and filtration. The mesenchymal stem cells of mouse fetus and mouse placenta were isolated with density gradient centrifugation, two steps centrifugalization and adhesive screening methods. The cells were identified with cell-surface markers and cultured in an adipogenic medium or in an osteogenic medium. After induction, the cells were identified with oil red O staining and alizarin bordeaux staining respectively.2. Implantation of cells suspension and mesenchymal stem cells.Firstly,6-day-old fetus cells suspension and 18-day-old placenta tissue cells suspension were administrated respectively into abdominal cavity of 15-month-old naturally aging female BALB/c mice of different groups with intraperitoneal injection respectively.Secondly, the mesenchymal stem cells from them were implanted respectively into 15-month-old naturally aging female BALB/c mice of other different groups with tail vein injection in other time.3. Viewing objects before and after anti-aging treatmentThe mice survival time were recorded after anti-aging treatment. The mice health situation were evaluated with ultrasoundcardiogram on cardiac output, stroke volume, left ventricle end-diastolic volume and left ventricular mass before and 3 month later after anti-aging treatment. Firstly, the serum total superoxide dismutase activity, serum maleic dialdehyde content and serum glutathione peroxidase activity were detected after the second ultrasoundcardiogram. Secondly the mice were evaluated with heart mass index representing heart function (heart mass/body mass), spleen mass index representing immune function (spleen mass/body mass), and colon muscular layer thickness representing digestive tract dynamia, and we detected the oncogenicity of stem cells with anatomy and evaluated the mice with score of cardia, kidney, lung, skin histopathology at the same time. At last the imbedded cells were traced in organ tissues of died mice with in situ Y chromosomal hybridization staining.Results:1. The cell-surface marker of mouse fetus and placenta derived mesenchymal stem cells was in accordance with that of mesenchymal stem cells. After induction the cells were positive for oil red O staining and alizarin bordeaux staining.2. After implantation, the surviving time of mouse fetus tissue cells suspension group, mouse placenta tissue cells suspension group and mouse fetus derived mesenchymal stem cells group were at least 1.3 times of the corresponding control one, and the difference were significant (all P<0.05)3. After implantation, we did not find any harmful effect with the implantation, nor any macroscopic tumor with all of postmortal anatomy inspection, but found the long-term surviving implanted cells located in organ tissues of all implanted mice with in situ Y chromosomal hybridization staining.4. After implantation, many indexes showed the significant differences between mouse fetus tissue cells suspension implantation group, mouse placenta tissue cells suspension implantation group, mouse fetus derived mesenchymal stem cells implantation group and their corresponding control group, such as decrease of cardiac output, decrease of stroke volume, increase of left ventricular mass, decrease of left ventricle end-diastolic volume, heart mass index, spleen mass index, colon muscular layer thickness, the score of heart, kiney, lung, skin histopathology, serum total superoxide dismutase activity, maleic dialdehyde content and glutathione peroxidase activity (all P<0.05). 5. But there was no statistical significance on the difference of pre-stated indexes except for serum total superoxide dismutase activity, maleic dialdehyde content and glutathione peroxidase activity between mouse placenta derived mesenchymal stem cells implantation group and the control group.Conclusions:1. Mesenchymal stem cells can be isolated from mouse placenta.2. Both mouse fetus derived cell suspension and mesenchymal stem cells and mouse placenta derived mesenchymal stem cells all have anti-aging capacity in mouse.3. Mouse placenta derived mesenchymal stem cells has no conclusive anti-aging effect on mouse.