Iberian pig mesenchymal stem/stromal cells from dermal skin, abdominal and subcutaneous adipose tissues, and peripheral blood: in vitro characterization and migratory properties in inflammation

Abstract

BackgroundRecently, the capacity of mesenchymal stem/stromal cells (MSCs) to migrate into damaged tissues has been reported. For MSCs to be a promising tool for tissue engineering and cell and gene therapy, it is essential to know their migration ability according to their tissue of origin. However, little is known about the molecular mechanisms regulating porcine MSC chemotaxis. The aim of this study was to examine the migratory properties in an inflammatory environment of porcine MSC lines from different tissue origins: subcutaneous adipose tissue (SCA-MSCs), abdominal adipose tissue (AA-MSCs), dermal skin tissue (DS-MSCs) and peripheral blood (PB-MSCs).MethodsSCA-MSCs, AA-MSCs, DS-MSCs and PB-MSCs were isolated and analyzed in terms of morphological features, alkaline phosphatase activity, expression of cell surface and intracellular markers of pluripotency, proliferation, in vitro chondrogenic, osteogenic and adipogenic differentiation capacities, as well as their ability to migrate in response to inflammatory cytokines.ResultsSCA-MSCs, AA-MSCs, DS-MSCs and PB-MSCs were isolated and showed plastic adhesion with a fibroblast-like morphology. All MSC lines were positive for CD44, CD105, CD90 and vimentin, characteristic markers of MSCs. The cytokeratin marker was also detected in DS-MSCs. No expression of MHCII or CD34 was detected in any of the four types of MSC. In terms of pluripotency features, all MSC lines expressed POU5F1 and showed alkaline phosphatase activity. SCA-MSCs had a higher growth rate compared to the rest of the cell lines, while the AA-MSC cell line had a longer population doubling time. All MSC lines cultured under adipogenic, chondrogenic and osteogenic conditions showed differentiation capacity to the previously mentioned mesodermal lineages. All MSC lines showed migration ability in an agarose drop assay. DS-MSCs migrated greater distances than the rest of the cell lines both in nonstimulated conditions and in the presence of the inflammatory cytokines TNF-α and IL-1β. SCA-MSCs and DS-MSCs increased their migration capacity in the presence of IL-1β as compared to PBS control.ConclusionsThis study describes the isolation and characterization of porcine cell lines from different tissue origin, with clear MSC properties. We show for the first time a comparative study of the migration capacity induced by inflammatory mediators of porcine MSCs of different tissue origin.

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

“…MSCs in that study did not show an increase in CD31 expression, suggesting that they did not differentiate in alternative lineages up to passage 3 or 4 under EGM. We found similar surface marker phenotype profiles as other groups (11,27,28,30,33) and there was no relevant difference between the cells. CD73 is known for lack of cross-reactivity of the CD73 antibody in pigs and was confirmed here (16,34).…”

“…In a study by Toyoda et al, isolation of human ASCs from omentum majus and subcutis yielded fewer and more cells, respectively, compared to our study, probably due to the differing isolation protocol (12). As far it concerns pig-derived cells, Calle et al found that ASCs from abdominal fat have longer PDT than their SC counterpart under culture with DMEM (27), which is in contrast to similar PDT in O-ASC and SC-ASC in our study under EGM. Other reports showed that BM-MSC had higher PDT than ASCs in the past (28)(29)(30), which was confirmed only in the first passage in our study.…”

Bovine peripheral blood MSCs chemotax towards inflammation and embryo implantation stimuli

“…We measured the invasion capacity of all pbMSCs lines using the agarose spot assay (Calle et al, 2018(Calle et al, , 2019, which allows the as- To analyze the migration capacity of the pbMSC lines using cytokine stimulation, assays were performed at a shorter period (48 hr)…”

“…For adipogenic, osteogenic, and chondrogenic differentiation, samples were analyzed according to manufacturer's instructions of the StemPro® adipogenesis, osteogenesis, or chondrogenesis differentiation kits, respectively (Thermo Fisher Scientific, Meridian Road Rockford, IL) and as detailed inCalle et al (2018).…”

Differentiation Potential of Mesenchymal Stem/Stromal Cells Is Altered by Intrauterine Growth Restriction

“…These are important aspects that should be taken into consideration during selection of MSCs for regenerative therapy applications. Porcine MSCs have been characterized (47,48), and they display features, in terms of marker expression and multipotency, that are similar to those of companion animals and human MSCs (49). Here, we showed that, similar to Normal-MSCs, IUGR-MSCs displayed classical features of human and porcine MSCs (49-51) namely of CD marker expression, as defined by ISCT (9), and, except for a faster initial cell growth of Normal-MSCs, there were no differences between the two cell types.…”