Characterization of mesenchymal stem cells of "no-options" patients with critical limb ischemia treated by autologous bone marrow mononuclear cells

Cestmir Altaner; Veronika Altanerova; Marina Cihova; Lubica Hunakova; Katarina Kaiserova; Andrej Klepanec; Ivan Vulev; Juraj Madaric

doi:10.1371/journal.pone.0073722

Characterization of mesenchymal stem cells of "no-options" patients with critical limb ischemia treated by autologous bone marrow mononuclear cells

PLoS One. 2013 Sep 12;8(9):e73722. doi: 10.1371/journal.pone.0073722. eCollection 2013.

Authors

Cestmir Altaner¹, Veronika Altanerova, Marina Cihova, Lubica Hunakova, Katarina Kaiserova, Andrej Klepanec, Ivan Vulev, Juraj Madaric

Affiliation

¹ Cancer Research Institute, Slovak Academy of Sciences, Bratislava, Slovakia ; St. Elisabeth Cancer Institute, Bratislava, Slovakia.

Abstract

Background: Application of autologous bone marrow mononuclear cells to "no option" patients with advanced critical limb ischemia (CLI) prevented major limb amputation in 73% patients during the 6-month follow-up. We examined which properties of bone marrow stromal cells also known as bone-marrow derived mesenchymal stem cells of responding and non-responding patients are important for amputation-free survival.

Methods and findings: Mesenchymal stem cells of 41 patients with CLI unsuitable for revascularisation were isolated from mononuclear bone marrow concentrate used for their treatment. Based on the clinical outcome of the treatment, we divided patients into two groups: responders and non-responders. Biological properties of responders' and non-responders' mesenchymal stem cells were characterized according to their ability to multiply, to differentiate in vitro, quantitative expression of cell surface markers, secretion of 27 cytokines, chemokines and growth factors, and to the relative expression of 15 mesenchymal stem cells important genes. Secretome comparison between responders (n=27) and non-responders (n=14) revealed significantly higher secretion values of IL-4, IL-6 and MIP-1b in the group of responders. The expression of cell markers CD44 and CD90 in mesenchymal stem cells from responders was significantly higher compared to non-responders (p<0.01). The expression of mesenchymal stem cells surface markers that was analyzed in 22 patients did not differ between diabetic (n=13) and non-diabetic (n=9) patient groups. Statistically significant higher expression of E-cadherin and PDX-1/IPF1 genes was found in non-responders, while expression of Snail was higher in responders.

Conclusions: The quality of mesenchymal stem cells shown in the expression of cell surface markers, secreted factors and stem cell genes plays an important role in therapeutic outcome. Paracrine mechanisms are main drivers in the induction of reparatory processes in CLI patients. Differences in mesenchymal stem cells properties are discussed in relation to their involvement in the reparatory process.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aged
Bone Marrow Cells / cytology*
Cells, Cultured
Female
Flow Cytometry
Humans
Ischemia / therapy*
Leukocytes, Mononuclear / cytology*
Male
Mesenchymal Stem Cells / cytology*
Middle Aged

Grants and funding

Laboratory part of the study was supported by grants from the Slovak League against Cancer, and VEGA - Grant Funding Agency of the Slovak Academy of Sciences (2/0177/11). Clinical study was sponsored by a grant from European Regional Development Funding (ITMS code: 26240220023). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.