Expression analysis of multiple myeloma CD138 negative progenitor cells using single molecule microarray readout

Jaroslaw Jacak, Harald Schnidar, Leila Muresan, Fritz Aberger, Gerhard Schütz, Jan Hesse

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

We present a highly sensitive bioanalytical microarray assay that enables the analysis of small genomic sample material. By combining an optimized cDNA purification step with single molecule cDNA detection on the microarray, the platform has improved sensitivity compared to conventional systems, allowing amplification-free determination of expression profiles with as little as 600ng total RNA. Total RNA from cells was reverse transcribed into fluorescently labeled cDNA and purified employing a precipitation method that minimizes loss of cDNA material. The microarray was scanned on a fluorescence chip-reader with single molecule sensitivity. Using the newly developed platform we were able to analyze the RNA expression profile of a subpopulation of rare multiple myeloma CD138 negative progenitor (MM CD138neg) cells. The high-sensitivity microarray approach led to the identification of a set of 20 genes differentially expressed in MM CD138neg cells. Our work demonstrates the applicability of a straight-forward single-molecule DNA array technology to current topics of molecular and cellular cancer research, which are otherwise difficult to address due to the limited amount of sample material.

Original languageEnglish
Pages (from-to)525-530
Number of pages6
JournalJournal of Biotechnology
Volume164
Issue number4
DOIs
Publication statusPublished - 5 Apr 2013

Keywords

  • Cell Line, Tumor
  • Gene Expression Profiling/methods
  • Humans
  • Microscopy, Fluorescence/methods
  • Multiple Myeloma/metabolism
  • Neoplastic Stem Cells/chemistry
  • Oligonucleotide Array Sequence Analysis/methods
  • RNA, Messenger/analysis
  • Stem Cells/chemistry
  • Syndecan-1/genetics
  • Expression profiling
  • Single molecule detection
  • Subpopulation analysis
  • Microarray
  • Fluorescence microscopy

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