Priming mobilized peripheral blood mononuclear cells with the "activated platelet supernatant" enhances the efficacy of cell therapy for myocardial infarction of rats

Cardiovasc Ther. 2016 Aug;34(4):245-53. doi: 10.1111/1755-5922.12194.

Abstract

Aim: Various methods are used to augment the efficacy of cell therapy in myocardial infarction (MI). In this study, we used the "activated platelet supernatant (APS)" to prime autologous "granulocyte colony-stimulating factor-mobilized peripheral blood mononuclear cells ((mob) PBMCs)" and investigated the efficacy of cell-based therapy in MI.

Method: Rat (mob) PBMCs were isolated after daily subcutaneous injections of G-CSF at 100 μg/kg for 3 days. APS was isolated separately after activating rat platelets with thrombin 0.5 U/mL for 2 hours. Priming was performed with APS for 6 hours. To check the paracrine effect of primed (mob) PBMCs, we used the 36-hour culture supernatant of the primed cells. A rat MI model was used for an in vivo model.

Result: Cytokines such as IL-1β, IL-10, and TGFβ were 3.7±0.9-fold, 3.4±1.2-fold, and 1.2±0.1-fold higher in APS, respectively, compared with naïve platelet supernatant. By APS priming, (mob) PBMCs showed M2 polarization and upregulation of angiogenic molecules (i.e., TEK, IL-10, CXCL1, and CX3CR1). APS-primed (mob) PBMCs had a 2.3-fold increased adhesion ability, induced by upregulated integrins. Rat endothelial cells cultured in the 36-hour culture supernatant of APS-primed (mob) PBMCs showed a 1.6-fold augmented proliferation and capillary network formation. In vivo transplantation of APS-primed (mob) PBMCs into rat MI models showed a significant trend of reduction in fibrosis area (P=.001) and wall thinning (P=.030), which lead to improvement in cardiac function measured by echocardiography.

Conclusion: Our data reveal that APS priming can enhance the wound-healing potential of (mob) PBMCs. APS priming may be a promising method for cell-based therapy of MI.

Keywords: Activated platelet supernatant; Cell Therapy; Cell priming; Granulocyte colony-stimulating factor-mobilized peripheral blood mononuclear cells; M2 polarization; Myocardial infarction.

MeSH terms

  • Angiogenic Proteins / metabolism
  • Animals
  • Blood Platelets / metabolism*
  • Cell Adhesion Molecules / metabolism
  • Cell Proliferation
  • Cells, Cultured
  • Cytokines / blood
  • Disease Models, Animal
  • Endothelial Cells / metabolism
  • Granulocyte Colony-Stimulating Factor / administration & dosage
  • Injections, Subcutaneous
  • Leukocytes, Mononuclear / drug effects
  • Leukocytes, Mononuclear / metabolism
  • Leukocytes, Mononuclear / transplantation*
  • Myocardial Infarction / blood
  • Myocardial Infarction / pathology
  • Myocardial Infarction / physiopathology
  • Myocardial Infarction / surgery*
  • Neovascularization, Physiologic
  • Paracrine Communication*
  • Phenotype
  • Platelet Activation*
  • Rats, Sprague-Dawley
  • Recovery of Function
  • Subcellular Fractions / metabolism
  • Time Factors
  • Wound Healing

Substances

  • Angiogenic Proteins
  • Cell Adhesion Molecules
  • Cytokines
  • Granulocyte Colony-Stimulating Factor