Proteomic Analysis of Chorion-Derived Mesenchymal Stem Cells: Combination of 2D Nano-HPLC in Tandem with ESI Mass Spectrometry
Abstrakt
The human placenta plays an important role in foetus development and growth and is also well known as the reservoir of the mesenchymal stem cells (MSCs), which are characterized by significantly higher plasticity in comparison to adult stem cells as well as by the ability to differentiate into multiple cell types. Therefore, they are promising candidates for stem-cell therapy.
To understand that mechanism the description of their proteomic profile is required. In this context, the goal of the present study was to isolate and analyse the proteome from chorionic membrane of the human placenta.
The whole-cell protein extracts were analysed by LC MS/MS utilizing bottom-up proteomic approach. To maximize the number of identified proteins, two different nanoLC approaches were used.
In the first approach only one-dimensional pre-concentration setup (C-18 reverse-phase column) was used; whereas, the second approach utilized the two-dimensional LC in a salt plug setup (first dimension-SCX column and second dimension-C-18 reverse-phase column). The method validation was accomplished using foetal membranes of six donors.
Identification of 334, respectively, 500 proteins as part of chorionic MSCs proteome is reported here, supported by gene ontology classification and systematic protein family ordering. Complete characterization of chorionic stem-cell proteome profile will contribute to identification and understanding of the molecular pathways involved in the cell processes such as self-renewal, proliferation and differentiation.