The Study Of Effect Of Discontinuous Pressure To Adulte's MSC_s Cultured In Vitro

Objective: The main cause of the bone tissue engineering haven' t been apph'cated extensively in clinical is the datebases of laboratory is sufficient, staying in animal' s experiment stage mostly. We quote the achievements of predecessor to culture adulte' s MSCs in vitro under discontinuous pressure, investigating the relation of cell' s biological properties and discontinuous pressure, to accelerate bone tissue engineering to applicate in clinical.Methods: To isolate and purify MSCs from health adultes' marrow and divide into 3 groups. (1) group of detection: inoculate cells in 6 pores with inductive factors or routine culture medium ,to observe two groups' cells integrity form and ultrastructure with invert microscope, HE stain, electron microscope, and to decisive quality with ALP stain. calcium nodes stain. (2 ) group of primary culture under pressure: inoculate cells in 96 pores, tese group-to seal the plates and culture under discontinuous pressure(staying in 0.098Mpa for 15 miniates and OMpa for 15 miniates,8 hours per day,)after 72 hours,continuing for 9 days. Blank group-all conditions are alike to the test group except to no pressure.then to detect the 2 groups' cells'quantity and ALP activity at the 6th , 9th, 12th day after inoculation. (3) group of subcultule under pressure:when we found cells proliferation no more change and no ALP express after routine primary culture after several times passage, putting cells under the same preseeure as the group of primary culture under pressure ,then detect the ALP again.Results: (1) MSCs can converse into osteoblasts spontaneously or by induction.inductive factors accelarate differentiation but inaccelarate proliferation. (2) The cells' differentiation and proliferation of test group is increasing comparedwith the blank group, and the difference is significant.But as the time going ,two groups' speed has no difference. (P<0.05)(3) We put the MSCs which have no many changes in proliferation and differentiation under discontinuous pressure and detect ALP again, finding there are no significant changes.Conclusion: (1)Human' s MSCs are immatural stem cells which can differentiate into osteoblasts by isolating and purificating in vitro.(2)Reasonable cultivation and external expression can accalerate proliferation and differentiation.(3) Discontinuous pressure is no significant use in proliferation and differentiation to senile MSCs.