Isolation and characterization of multipotent mesenchymal stem cells in nasal polyps

Abstract

Mesenchymal stem cells (MSCs) are multipotent progenitor cells in adult tissues. This study aimed to investigate nasal polyp (NP) tissues as a potential new source of multipotent MSCs that maintain their stemness and differentiation potential following multiple rounds of passaging. NP tissues were obtained from 10 patients during endoscopic sinus surgery. After isolating and culturing NPderived MSCs (npMSCs), the expression levels of the surface markers CD34, CD44, CD45, CD73, CD90, CD105, CD106, CD146 and human leukocyte antigens-class II DR antigen (HLA-DR) were estimated by flow cytometry. NpMSCs were cultured in chondrogenic, osteogenic, adipogenic, or neurogenic differentiation medium. The differentiation potential of npMSCs was analyzed by Alcian blue, alizarin red S, oil red O, and immunocytochemical staining and reverse transcription-polymerase chain reaction. The clonogenic potential of npMSCs was measured using a colony-forming unit assay. Cell proliferation of npMSCs was measured using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay. Flow cytometry analysis revealed that npMSCs were negative for hematopoietic lineage markers (CD34, CD45, and HLA-DR) and positive for MSC markers (CD44, CD73, CD90, and CD105). The npMSCs differentiated into osteogenic, adipogenic, chondrogenic, and neurogenic lineages, respectively. Chondrogenically differentiated npMSCs were stained with Alcian blue, osteogenically differentiated npMSCs were stained with alizarin red S, and adipogenically differentiated npMSCs were stained with oil red O. Real-time polymerase chain reaction results showed that the differentiated npMSCs expressed the respective differentiation markers (Sox 9 and Col2A for chondrogenesis, Runx2 and osteocalcin for osteogenesis, fatty acid-binding protein 4 and peroxisome proliferator-activated receptor for adipogenesis, TuJ1, neurofilament light chain, and neurofilament heavy chain for neurogenesis). There were no significant differences in the clonogenic potential and proliferation rate between early and late passage npMSCs. These results show that npMSCs possess the characteristics of MSCs in terms of morphology, multipotent differentiation capacity, cell surface marker expression, and clonogenicity. Thus, npMSCs may represent an alternative source of MSCs.

Neuronal Differentiation Capability of Nasal Polyps of Chronic Rhinosinusitis

“…Until now, only a few researchers have addressed this issue. Cho et al (2015) isolated and characterized MSC in nasal polyps and proposed nasal polyp-derived MSCs as an alternative source of MSCs. Otherwise, Pezato et al (2014) demonstrated that bone marrow-derived MSCs had an immunoregulatory effect on immune cells (especially T cells) derived from nasal tissue affected by CRSwNP.…”

“…In detail, researchers linked the pathogenesis of nasal polyps to the presence and differentiation of mesenchymal stem cells (MSC) (Cho et al 2014(Cho et al , 2015Pezato et al 2014), pathological changes in the epithelium (de Borja Callejas et al 2014;Li et al 2014;Wiszniewski et al 2006;Yu et al 2014) or to single stem cell markers like stem cell factor, Nestin, and BMI-1 (Kim et al 1997(Kim et al , 2009Kowalski et al 2005). The overall stemness and for example neuronal differentiation potential of nasal polyps are still unknown.…”

Cardiac Analysis of Autologous Transplantation of Cocultured Skeletal Myoblasts and Mesenchymal Cells in a Rat Model Doxorubicin-Induced Cardiotoxicity: Histopathological and Functional Studies

“…Flow cytometric analyses were employed to mesenchymal stem cell characterization by CD73, CD90, and CD105 positive cell surface marker expression, while being negative for CD34, CD45, and HLA-DR surface markers [12,22].…”

“…MSCs were washed with phosphate buffered solution (pH 7.4) and incubated in the dark for 1 h at room temperature with CD45-FITC or CD34-PE-Cy7 antibodies (BD Pharmingen, San Diego, CA, USA), following the protocol previously described [12]. The specific fluorescence of 10,000 cells was analyzed on FACScalibur (Becton Dickinson, Franklin Lakes, NJ, USA) using Cell Quest Pro software.…”

PPAR Agonists Promote the Differentiation of Porcine Bone Marrow Mesenchymal Stem Cells into the Adipogenic and Myogenic Lineages

“…They are characterized by a high rate of renewal, proliferative ability and multipotentiality. The latter term refers to their ability to differentiate into various cell lineages, such as chondrocytes [Mathieu et al, 2014], osteocytes [Cho et al, 2014], adipocytes [Casado et al, 2012], neurons [Nandy et al, 2014], and myocytes [Meligy et al, 2012].…”