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Characteristics between porcine bone marrow-derived mesenchymal stem and peripheral blood mononuclear cells

ABSTRACT: Mesenchymal stem cells (MSCs) have been isolated from various organs and extensively studied for their potential in regulating transplantation. MSCs from different mammalian species are well characterized; however, the properties and therapeutic potential of porcine bone marrow-derived MSCs (BM-MSCs) remain unclear. In this study, we aimed to profile the characteristics of porcine BM-MSCs by comparing their gene expression patterns and immunomodulatory properties with those of porcine peripheral blood mononuclear cells (PBMCs) and bone marrow-attached cells (BMACs). Using quantitative polymerase chain reaction, flow cytometry, immunocytochemistry, and RNA sequencing, we confirmed the expression of key MSC markers, including CD105, CD73, and CD90, in porcine BM-MSCs, and aligned them closely with human MSCs. We found significant differences in gene expression between BM-MSCs and PBMCs, with BM-MSCs exhibiting a distinct expression pattern similar to that of BMACs. Gene ontology enrichment analysis revealed the pathways involved in immune modulation and tissue repair, underscoring the potential of BM-MSCs to enhance immune regulation. Notably, BM-MSCs exhibited higher transforming growth factor-beta levels than PBMCs, suggesting a central role in their immunosuppressive function. These findings indicate the immunomodulatory capabilities of porcine BM-MSCs and support their application in xenotransplantation, where they may help mitigate graft rejection and promote tissue regeneration.

ORGANISM(S): Sus scrofa

PROVIDER: GSE284117 | GEO | 2025/03/31

REPOSITORIES: GEO

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Project description:Objective: Craniofacial bone defects caused by injuries and congenital diseases are a formidable challenge to clinicians. Research has shown promise in using bone marrow mesenchymal stem cells (BM-MSCs) from limb bones for craniofacial bone regeneration; yet little is known about the potential of BM-MSCs from craniofacial bones. This study compared BM-MSCs isolated from limb and craniofacial bones in pigs, a preclinical model closely resembling humans. Design: Bone marrow was aspirated from the tibia and mandible of four-month-old pigs (n=4), followed by BM-MSC isolation, culture-expansion and confirmation by flow cytometry. Proliferation rates were compared using population doubling times. Osteogenic differentiation was evaluated by quantifying alkaline phosphatase (ALP) activity. Total mRNA was extracted from freshly isolated BM-MSCs and analyzed to compare gene expressions of tibial and mandibular BM-MSCs using an Affymetrix GeneChip porcine genome array, followed by real-time RT-PCR evaluation of two neural crest markers. Results: BM-MSCs from both locations expressed MSC markers without expression of hematopoietic markers. Mandibular BM-MSCs proliferated significantly faster than tibial BM-MSCs. Without osteogenic inducers, mandibular BM-MSC alkaline phosphatase activities were 3.3-fold greater than those of tibial origin. Microarray analysis identified 383 differentially expressed genes in mandibular and tibial BM-MSCs, including higher expression of cranial neural crest-related genes nestin and BMP-4 in mandibular BM-MSCs, a trend also confirmed by real-time RT-PCR. Among differently expressed genes, only 47 showed greater than 1.5-fold differences in expression. Conclusions: These data indicate that despite many similarities in gene expression, mandibular BM-MSCs express of number of genes differently than tibial BM-MSCs and have a phenotypic profile that may make them advantageous for craniofacial bone regeneration. Bone marrow was aspirated from the mandibular symphyseal region and the tibia of 3 pigs. Mesenchymal stem cells were isolated from the bone marrow and cultured to 80% confluence. Cells were harvested for total RNA extraction and the RNA was analyzed by Affymetrix GeneChip porcine genome array.

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Characteristics between porcine bone marrow-derived mesenchymal stem and peripheral blood mononuclear cells

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