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Bone & Joint Research
Vol. 5, Issue 11 | Pages 569 - 576
1 Nov 2016
Akahane M Shimizu T Kira T Onishi T Uchihara Y Imamura T Tanaka Y

Objectives

To assess the structure and extracellular matrix molecule expression of osteogenic cell sheets created via culture in medium with both dexamethasone (Dex) and ascorbic acid phosphate (AscP) compared either Dex or AscP alone.

Methods

Osteogenic cell sheets were prepared by culturing rat bone marrow stromal cells in a minimal essential medium (MEM), MEM with AscP, MEM with Dex, and MEM with Dex and AscP (Dex/AscP). The cell number and messenger (m)RNA expression were assessed in vitro, and the appearance of the cell sheets was observed after mechanical retrieval using a scraper. β-tricalcium phosphate (β-TCP) was then wrapped with the cell sheets from the four different groups and subcutaneously implanted into rats.


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XIV | Pages 18 - 18
1 Apr 2012
Miyachi M Yoshida H Fujiki A Yagyu S Kikuchi K Tsuchiya K Imamura T Iehara T Hosoi H
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Aim

The recent discovery of small non-coding RNAs, so-called micro RNAs (miRNAs), has provided new insights into cancer diagnosis. Several studies have shown that profiles of miRNA expression differ between normal tissue and tumour tissue and vary among tumour types. To exploit this difference, we evaluated the feasibility of using muscle-specific miRNAs (miR-1, 133a, 133b, 206) as biomarkers of rhabdomyosarcoma (RMS).

Method

Total RNA was extracted from 16 cell lines (7 RMS, 4 neuroblastoma, 3 Ewing sarcoma and 2 malignant rhabdoid tumour) and 21 tumour specimens (7 RMS, 1 Ewing sarcoma, 4 undifferentiated sarcoma, 1 osteosarcoma, 1 alveolar soft part sarcoma, 2 neuroblastoma, 2 Wilms tumour, 1 malignant rhabdoid tumour, 1 adrenal carcinoma and 1 retinoblastoma). miRNA was quantified by real-time RT-PCR. The expression levels of miRNAs were calculated utilizing the delta-delta Ct method, normalised to the level of miR-16, and compared using the Mann-Whitney U test.