Evaluation of Porcine Versus Human Mesenchymal Stromal Cells From Three Distinct Donor Locations for Cytotherapy

Abstract

Background: Mesenchymal stromal cell (MSC)-based cytotherapies fuel the hope for reduction of chronic systemic immunosuppression in allotransplantation, and our group has previously shown this capability for both swine and human cells. MSCs harvested from distinct anatomical locations may have different behavior and lead to different outcomes in both preclinical research and human trials. To provide an effective reference for cell therapy studies, we compared human and porcine MSCs from omental fat (O-ASC), subcutaneous fat (SC-ASC) and bone marrow (BM-MSC) under rapid culture expansion with endothelial growth medium (EGM). Methods: MSCs isolated from pigs and deceased human organ donors were compared for yield, viability, cell size, population doubling times (PDT), surface marker expression and differentiation potential after rapid expansion with EGM. Immunosuppressant toxicity on MSCs was investigated in vitro for four different standard immunosuppressive drugs. Immunomodulatory function was compared in mixed lymphocyte reaction assays (MLR) with/without immunosuppressive drug influence. Results: Human and porcine omental fat yielded significantly higher cell numbers than subcutaneous fat. Initial PDT was significantly shorter in ASCs than BM-MSCs and similar thereafter. Viability was reduced in BM-MSCs. Porcine MSCs were positive for CD29, CD44, CD90, while human MSCs expressed CD73, CD90 and CD105. All demonstrated confirmed adipogenic differentiation capacity. Cell sizes were comparable between groups and were slightly larger in human cells. Rapamycin revealed slight, mycophenolic acid strong and significant dose-dependent toxicity on viability/proliferation of almost all MSCs at therapeutic concentrations. No relevant toxicity was found for Tacrolimus and Cyclosporin A. Immunomodulatory function was dose-dependent and similar between groups. Immunosuppressants had no significant adverse effect on MSC immunomodulatory function. Schweizer et al. MSCs From Distinct Donor Locations Discussion: MSCs from different harvest locations and donor species differ in terms of isolation yields, viability, PDT, and size. We did not detect relevant differences in immunomodulatory function with or without the presence of immunosuppressants. Human and pig O-ASC, SC-ASC and BM-MSC share similar immunomodulatory function in vitro and warrant confirmation in large animal studies. These findings should be considered in preclinical and clinical MSC applications.

Morphological and Molecular Features of Porcine Mesenchymal Stem Cells Derived From Different Types of Synovial Membrane, and Genetic Background of Cell Donors

“…Beyond the consideration of the use of procine tissues as excellent large animal model for human in the development of stem cell based therapies, regenerative medicine and transplantation for preclinical research, the generation of porcine MSCs will establish background knowledge and technology in a variety of experimental research for both veterinary clinicians and livestock industry ( Swindle et al, 2012 ; Arrizabalagaa and Nollerta, 2017 ; Bharti et al, 2016 ; Markoski, 2016 ; Schweizer et al, 2020 ). Application of MSCs may be a powerful tool for treatment of several animal health conditions, and some of the major skeletal abnormalities and disorders that can result in loss of both animal welfare and economic benefits ( Fan et al, 2009 ; Ogata et al, 2015 ; Gugjoo et al, 2019 ).…”

Culture and characterization of various porcine integumentary-connective tissue-derived mesenchymal stromal cells to facilitate tissue adhesion to percutaneous metal implants

“…1 b). The expression of CD105 as a pMSC marker in pigs is still controversial [ 24 , 25 ]. The pattern of gene transcript expression for the pMSC-specific markers varied across the nine tissues; molar-derived periodontal ligament (Molar), bone marrow and hind limb (HL) dermis-derived cells expressed significantly higher levels of the CD90 transcript, compared to the other cell types; while abdominal (Ab) dermis, hind limb adipose and tendon-derived cells expressed significantly high levels of the CD44 transcript.…”

“…Tissue-specificity and selectivity of the nine pMSCs was also reflected in the differentiation studies, with distinct transcription factors upregulated in the different pMSCs during the early stages of differentiation. It is important to consider species-specific differences between porcine and human MSCs that came up in this study, exemplified by the dim expression of CD105 on pMSCs, which on the other hand, is a well-recognized marker for human MSCs [ 23 , 25 ]. Others have reported CD105 is [ 42 ] and is not [ 25 ] expressed by pMSCs, further supporting tissue-specificity.…”

“…It is important to consider species-specific differences between porcine and human MSCs that came up in this study, exemplified by the dim expression of CD105 on pMSCs, which on the other hand, is a well-recognized marker for human MSCs [ 23 , 25 ]. Others have reported CD105 is [ 42 ] and is not [ 25 ] expressed by pMSCs, further supporting tissue-specificity. Here, we were able to detect low levels of mRNA gene transcript and cell surface protein expression of CD105.…”

Identifying Key Genes and Functionally Enriched Pathways of Diverse Adipose Tissue Types in Cattle

“…Adipose tissues from different anatomical locations usually differ in gene expression and regulatory mechanisms ( Schweizer et al, 2020 ; Silva and Baptista, 2019 ). According to the anatomical location, subcutaneous adipose tissues also vary in their gene expression and function ( Martos-Moreno et al, 2013 ).…”