Proteomic analysis of the extracellular matrix produced by mesenchymal stromal cells: implications for cell therapy mechanism

PLoS One. 2013 Nov 14;8(11):e79283. doi: 10.1371/journal.pone.0079283. eCollection 2013.


Mesenchymal stromal cells (MSCs) transiently transfected with notch1 intracellular domain (NICD) are beneficial for neurological disorders as observed in several preclinical studies. Extracellular matrix (ECM) derived from NICD-transfected MSCs has been previously shown to support in vitro neural cell growth and survival better than that of un-transfected MSCs. To understand the underlying mechanism(s) by which NICD-transfected MSC-derived ECM supports neural cell growth and survival, we investigated the differences in NICD-transfected MSC- and MSC-derived ECM protein quantity and composition. To compare the ECM derived from MSCs and NICD-transfected MSCs, the proteins were sequentially solubilized using sodium dodecyl sulfate (SDS) and urea, quantified, and compared across four human donors. We then analyzed ECM proteins using either in-gel digests or in-solution surfactant-assisted trypsin digests (SAISD) coupled with reverse phase nano-liquid chromatography and tandem mass spectrometry (nLC-MS/MS). Analyses using nLC-MS/MS identified key components of ECM from NICD-transfected MSCs and un-transfected MSCs and revealed significant differences in their respective compositions. This work provides a reproducible method for identifying and comparing in vitro cell-derived ECM proteins, which is crucial for exploring the mechanisms underlying cellular therapy.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cell Line
  • Cell- and Tissue-Based Therapy
  • Extracellular Matrix / genetics
  • Extracellular Matrix / metabolism*
  • Extracellular Matrix Proteins / genetics
  • Extracellular Matrix Proteins / metabolism
  • Gene Expression
  • Humans
  • Mesenchymal Stem Cells / metabolism*
  • Proteome*
  • Proteomics* / methods


  • Extracellular Matrix Proteins
  • Proteome

Grant support

Funding for this research project was provided by SanBio Inc. (AH, IA, CT, CC), the National Science Foundation Science Master’s Program Award DGE-1011717 (AH) and the National Science Foundation grant CHE- 0619163 (TY). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.