Nucleic Acid Sample Preparation from Stem Cells

Abstract

© Springer Science+Business Media New York 2016. The goal of this review is to give an integral overview on fundamental research in the stem-cell field, with emphasis on preparation for nucleic acid extraction out of these cells. Stem cells (SCs) have unlimited self-renewal, and large proliferative capacity, as well as a high potential for differentiation into all blood cells or other somatic cell types, in natural setting in situ and after “conventional” therapeutic use. In adult tissues they are dormant and can be mobilized by conventional mobilizers into peripheral blood, where they can be collected from, concentrated, and used for nucleic acid extraction, purified and analyzed dependent on the purpose. The most important nucleic acids within stem cells are: DNA (in the nucleus), RNA (in the cytoplasm), and miRNA (in the nucleus). There are few different techniques for preparation of the sample for nucleic acid extraction and purifi cation approach. Each has advantages and disadvantages. Thus, Fred Sanger received the Nobel Prize for Chemistry twice: in 1958 for sequencing the protein insulin and in 1980 for his contribution to the sequencing of DNA. Sample preparation—preparing nucleic acids or proteins for analysis—played a vital role in both achievements. It is obvious that without effective sample preparation, there is a risk that incorrect conclusions will be drawn. For example, important mRNA transcripts that have been degraded along the way will be missed, and damaged enzymes will give incorrect kinetics data. On the other hand, different types of stem cells require the most appropriate extraction. Therefore, the distinctive features of these types of cells are specified in this chapter with respect to different methods for nucleic acid extraction.