Abstract
Stem cells for regenerative medicine purposes offer therapeutic benefits, but disadvantages are still ill defined. The benefit of stem cells may be attributed to their secretion of growth factors (GFs), cytokines (CKs), and extracellular vesicles (EVs), including exosomes. We present a novel cell-free stem cell-derived extract (CCM), formulated from human progenitor endothelial stem cells (hPESCs), characterized for biologically active factors using ELISA, nanoparticle tracking analysis and single particle interferometric reflectance imaging sensing. The effect on fibroblast proliferation and ability to induce stem cell migration was analyzed using Alamar Blue proliferation and Transwell migration assays, respectively. GFs including IGFBP 1, 2, 3, and 6, insulin, growth hormone, PDGF-AA, TGF-α, TGF-β1, VEGF, and the anti-inflammatory cytokine, IL-1RA were detected. Membrane enclosed particles within exosome size range and expressing exosome tetraspanins CD81 and CD9 were identified. CCM significantly increased cell proliferation and induced stem cell migration. Analysis of CCM revealed presence of GFs, CKs, and EVs, including exosomes. The presence of multiple factors including exosomes within one formulation, the ability to promote cell proliferation and induce stem cell migration may reduce inflammation and pain, and augment tissue repair.
Highlights
During the past few decades, there has been a tremendous growth in the use of biologics for regenerative medicine applications [1,2]
Interleukin 1 receptor antagonist (IL-1RA), an anti-inflammatory cytokine was identified at significant levels in CCM relative to the control which had undetectable IL-1RA levels (Figure 1)
We describe the process of formulation of a novel cell-free stem cell-derived extract (CCM), and evaluated it for the presence of growth factors (GFs), CKs, and extracellular vesicles (EVs), including exosomes
Summary
During the past few decades, there has been a tremendous growth in the use of biologics for regenerative medicine applications [1,2]. Biologics currently available for clinical use include platelet rich plasma, bone marrow aspirate, lipoaspirate, amniotic allograft suspension, umbilical cord-derived Wharton’s Jelly, cord blood, and exosomes [3,4]. The efficacy of these biologics is attributed to the presence of stem cells, growth factors (GFs), cytokines (CKs), and extracellular vesicles (EVs), including exosomes [5,6]. MSCs present several disadvantages, including establishing a reliable source with stable phenotype, genetic instability and chromosomal aberrations, intravenous administration-related toxicity caused by physical trapping of the cells in the lung microvasculature, rejection by the host, formation of ectopic tissue, and tumorigenicity [11,12,13]
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