Telemere length and telomerase activity in cultured amniotic fluid cell

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Amniotic fluid cells are cultured in the condition of 37°C and 5% CO_2 using AmnioMax initially. Amniotic fluid cells were passed by 1st, 2nd, 5th respectively. In the passage 0.25% trypsin and MEM+10% FBS were used. The number of cells was treated 1X10^4.The pellet was reserved to measure telomerase activity at-80°C. We prepared 6 amniotic fluid cell samples, 3 chorionic villi samples and 1 fetus skin fibroblast.We used HeLa cells which have strong activity to perform the quantitative analysis about telomerase activity and made a measurement curve. We prepared 2 X10^4 cell count for HeLa cells and diluted cells by 5 steps respectively. We analyzed telomerase activity by using TeloChaser. The cell extract containing telomerase was prepared and telomere repeat sequence was added to substrate primer. After purification PCR was performed using reverse primer added. PCR products were analyzed by electrophoresis staining with ethidium bromide and a measurement curve was completed.The telomerase activity about amniotic fluid cells was analyzed using the same as above mentioned method. 6 amniotic fluid cells samples with normal chromosome karyotype had no telomerase activity.