Abstract:

BACKGROUND: Mid-half amniotic fluid was usually collected for mesenchymal stem cells (MSCs) culture, but the study on the other phase amniotic fluid-MSCs was seldom. The culture effect of amniotic fluid from different duration of pregnancy with different media is not reported.
OBJECTIVE: To compare the in vitro culture effect of MSCs in the amniotic fluid from different duration of pregnancy.
METHODS: Two media were used to isolate and culture MSCs from 60 cases amniotic fluid of pregnancy 15-42 weeks. The growth state and the success rate of MSCs cultivation course were observed. The cytochemical staining was applied to detect the chemical characteristics of MSCs. Growth curve of MSCs after passage was detected by MTT assay.
RESULTS AND CONCLUSION: The success rates of 37-42 weeks pregnancy amniotic fluid MSCs were 8% and 44% using PRMI-1640 and AmnioMAXⅡ complete medium containing 10% fetal bovine serum, that of 21-36 weeks were 20% and 60%; but that of 15-20 weeks was 100% using any medium. The result of cytochemical staining showed that POX and SB were negative; PAS, ACP, AENE were positive; NAP was 1% positive. No difference was determined in staining results at different duration of pregnancy. MTT assay showed that growth curve of MSCs exhibited “S” shape. Amniotic fluid-MSCs of 15-36 week pregnancy were transferred for 15 passages, showing prosperity; that of 37-42 weeks could only be transferred for 10 passages, showing slow growth. The results demonstrated that AmnioMAXⅡ complete medium can elevate obviously the success rate of amniotic fluid-MSCs from late-phase amniotic fluid. The optimal collection time of amniotic fluid was 15-20 weeks.