Human Term Placenta Mesenchymal Stem Cells In Vitro Purification And Biological Characteristics Of The Study

Objective:In order to search the new resource of Mecenchymal stem cells (MSCs),we investigated the human term placenta,isolated and purified MSCs from it. Then amplified and observed its biological characteristcs in vito, study the multitude potency of differentiating into adipose cells by oriented induction of human term placenta-drived marrow mesenchymal stem cells(hpMSCs). Methods:1.Human placenta mesenchymal stem cells were separated with four different methods in order to isolate MSCs:group A (Trypsin+Ficoll-hypaque method), group B (Trypsin/collagenaseⅣ+Hespan method), group C (collagenaseⅣ+Ficoll-hypaque method), group D (collagenaseⅣ+Hespan method). Five samples were in each group.2. The 3rd generation cells were identificated with immunofluorescence methods such as CD44 antibody, CD34 antibody, CD29 antibody,CD106 antibody; Observe biological properties of the cell with growth curve and electron microscopy.3. The 3rd generation cell were induced into adipose cells using containing isobutyl-methylxanthine, dexamethasone and trypsin, observe morphological changes of MSCs with oil red O staining, the group was not induced as a parallel control group. Result:1.Under the same conditions, the successful culture rate was 100% in group D, and 20%,60% and 80% in group A, B and C, respectively. The time of stretching and the time of primary culture with Hespan were shorter in the group D than that in other group.2.MSCs isolated from human term placenta were fibrous long spindle-like adherent cells and had strong proliferative ability;They were Strongly positive for CD44,CD29, but negative for CD34,CD106. 3.they also have the potential to differentiate into fat cells and the oil red O staining were positive. Conclusion: Collagenase IVdigestion combined with Hespan method could effectively extract mesenchymal stem cells from human placenta. MSCs isolated from human term placenta has the similar biological characteristics to mecenchymal stem cells from other sources. It was indicated that hpMSCs might be the new stem cells sources of tissue engineering.