| Project/Area Number |
17591141
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Embryonic/Neonatal medicine
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| Research Institution | Okayama University |
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Principal Investigator |
NINOMIYA Shinsuke Okayama University Hospital, Lecturer, 医学部・歯学部附属病院, 講師 (10325110)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Hiroyuki Okayama University Graduate School of Medicine and Dentistry, Associate Professor, 大学院医歯薬学総合研究科, 助教授 (80231413)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2006: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2005: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | amniotic fluid cell / chromosome / telomerase / テロメア長 / テロメラーゼ活性 |
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| Research Abstract |
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.
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