Preparation of thyroid tumor cells in aspiration biopsies for aspiration biopsy nucleic acid diagnosis

Abstract

Mesh filtration of aspirates proved superior results for the measurement of T/G ratio.

Trefoil Factor 3 (TFF3): A Promising Indicator for Diagnosing Thyroid Follicular Carcinoma

“…We reported a method to separate thyroid tumor cells from blood cells using mesh filtration (Fig. 2) [46]. After mesh filtration, the expression level of TFF3 mRNA in aspirates correlated well with that of corresponding tumor tissues.…”

“…Measurement of the expression level of TFF3 mRNA in aspirates is also problematic because of interference by contaminating peripheral blood cells or infiltrating lymphocytes. The expression level of TFF3 mRNA in aspirates does not always correlate with that in the corresponding tumor tissue [46]. We reported a method to separate thyroid tumor cells from blood cells using mesh filtration (Fig.…”

Measurement of <i>TFF3</i> mRNA in aspirates from thyroid nodules using mesh filtration: The first clinical trial in 130 cases

“…In our previous study, we established a novel method to separate thyroid tumor cells from blood cells using mesh filtration [24]. After mesh filtration, T/G ratios in aspirates corresponded well to those of corresponding tumor tissues.…”

“…The conditions for TaqMan PCR were as follows: 95°C for 2 minutes and 40 cycles of 95°C for 15 seconds and 60°C for 1 minute. Recombinant pGEM Easy T-Vector (Promega, Tokyo, Japan) containing TFF3, LGALS3, TG, PTPRC, or THBS-1 cDNA was constructed by PCR-cloning with the same set of primers used in TaqMan PCR, and Thyroglobulin (TG, GenBank X05615), CD45 (PTPRC, GenBank Y00062), and TSP-1 (THBS-1, GenBank X14787) mRNAs were also measured to estimate the number of thyroid tumor cells, white blood cells, and fibroblasts in the aspirates, respectively [24,25]. Realtime quantitative polymerase chain reaction (TaqMan PCR) using the ABI PRISM 7900 Sequence Detection System (Applied Biosystems, Foster, CA, U.S.A.) and Platinum qPCR SuperMix-UDG (Invitrogen) was performed according to the manufacturer's protocol.…”

“…All tissues were frozen in liquid nitrogen immediately after resection. Total RNA was extracted according to the method described previously [24].…”