The Comparison Of Somatic Cells From Different Sources Induced Into Pluripotent Stem Cells (iPS)

ObjectiveBy inducing various sources of somatic cells into induced pluripotent stem (iPS)cells, counting and computing the rate of iPS-like colonies, we compared andanalyzed these cell’s efficiency of being induced in order to find a method that couldget more iPS cells in a high efficiency and provide a better technical platform forregenerative medicine research.Methods1. Somatic cells from four different sources was isolated and cultured includingbone marrow mesenchymal stem cells(BMMSC), adipose-derived stemcells(ADSC), mouse embryonic fibroblast(MEF) and mouse tail fibroblast(MTF). All cells were stably passaged.2. The E. coli DH5α which transformed the plasmids with3transcriptionfactors was amplified cultured and respectively. After extraction andpurification, the plasmids were utilized to transfect Plat-E cells to makingvirus. The concentrated virus was dispensed and storaged in-80℃freezer.3. Somatic cells from four different sources were layered into4wells of a6-well plate, respectively and infected with concentrated virus. The infectedcells were cultured overnight and changed into fresh medium for continuedcultureing with the medium refreshing every2-3days. Monitoring the cellsand iPS-like coloniess. Counting the number of iPS-like coloniess.Results1. The BMMSC-like cells, ADSC-like cells, MEF-like cells and MTF-like cellswere harvested.2. The rates of iPS-like colonies were as follows: BMMSC0.255±0.025‰;ADSC0.278±0.038‰; MEF0.118±0.022‰; MTF0.138±0.022‰. Conclusion1. Preparation technic of bone marrow mesenchymal stem cells (BMMSC) waspreliminary established and BMMSC-like cell lines were harvested.2. Preparation technic of adipose-derived stem cells (ADSC) was preliminaryestablished and ADSC-like cell lines were harvested.3. The mouse tail fibroblast (MTF) preparation technic was successfullyestablished and MTF cell lines were harvested.4. Technology of induced pluripotent stem cells (iPSC) was preliminaryestablished.5. The induction efficiency of BMMSC and ADSC is higher than MEF andMTF. BMMSC and ADSC seems easier to be induced.