| [Objective] This experiment abide by principles of tissue engineering, collected ADSCs in different periods of pregnancy to be identificated, and cultivated in vitro, then analyzed their activity in proliferation to examine whether good result could be acquired by using ADSCs in pregnancy as seed cells, and also is useful explore to biological characteristics of ADSCs.[Methods] (1) Isolating and culturing of ADSCs, using light microscope to observe shape and status of the cells; (2) According the methods of examining surface antigen and experimental induction of bone formation to identify ADSCs; (3) Using MTT method and Propidium iodide staining to test absorbance value and cell cycle of third generation ADSCs in different groups respectively, drawing cells growth curve, then Using statistical analysis to evaluate results of the tests, compared the differences in proliferation of the ADSCs in different groups.[Results] (1) The cells in each groups grew well, the pattern of the spindle-shaped cells like whirlpool, the number of adherent spindle cells in pregnancy groups are much more than non-pregnant group. (2) Result of the surface antigen has showed the cells had cultured were CD90+CD44-CD34-, and experimental induction of bone formation showed red calcium nodules were produced. (3) The test of MTT showed:discrepancy between non-pregnant ADSCs+P group and non-pregnant ADSCs group didn't has significant different (P>0.05), discrepancy between ADSCs in middle period of pregnancy group, ADSCs in perinatal period, non-pregnant ADSCs+ E2 group didn't have significant different (P>0.05); ADSCs in middle period of pregnancy group, ADSCs in perinatal period, non pregnant ADSCs+ E2 group compared with non-pregnant ADSCs group, discrepancy has significant different (P<0.05); (4) PI stains showed: ADSCs in middle period of pregnancy group, ADSCs in perinatal period, non-pregnant ADSCs+ E2 group compared with non-pregnant ADSCs group, the percentage of cells in G0/G1 which is in non-proliferative period has a decrease trend, and have significant difference(p<0.05); non-pregnant ADSCs+P group compared with non-pregnant ADSCs group, ADSCs in middle period of pregnancy group compared with ADSCs in perinatal period group, the percentage of cells in G0/G1 period didn't has an apparent change, and didn't have significant different (p>0.05)[Conclusions] The cells had been cultured primary were ADSCs, and grew well under vitro circumstance; the proliferative status of ADSCs from pregnancy rat is better than the non-pregnant one. ADSCs in middle period of pregnancy compared with the perinatal one, the proliferative status didn't has apparent different; 17β-Estradiol could promote the proliferation of ADSCs, and Progesterone didn't show this ability. |
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