Update on canine MSC markers

Tryfonidou, Marianna A.; Schumann, Susanne; Armeanu, Sorin; Harichandan, Abhishek; Sivasubramaniyan, K.; Mollenhauer, Jürgen; Bühring, Hans‐Jörg

doi:10.1002/cyto.a.22469

Cited by 8 publications

(8 citation statements)

References 12 publications

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“…Finally, Tryfonidou et al. () reported an extensive testing of human antibodies that could be used to define canine MSC populations. In this review, they described CD271 and the in‐house generated antibodies to MSCA‐1, CD56, CD349, CD276 and CD34.…”

Section: Adult Stem Cellsmentioning

confidence: 99%

Stem Cells and Regenerative Medicine in Domestic and Companion Animals: A Multispecies Perspective

Gonçalves¹,

Ambrósio²,

Piedrahita

2014

Reprod Domestic Animals

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ContentsSince their original isolation, the majority of the work on embryonic stem cells (ESC) has been carried out in mice. While the mouse is an outstanding model for basic research, it also has considerable limitations for translational work, especially in the area of regenerative medicine. This is due to a combination of factors that include physiological and size differences when compared to humans. In contrast, domestic animal species, such as swine, and companion animal species, such as dogs, provide unique opportunities to develop regenerative medicine protocols that can then be utilized in humans. Unfortunately, at present, the state of knowledge related to, and availability of, ESC from domestic animals vary among species such as pig, horse, dog and cat, and without exception lags significantly behind the mouse and human. It is clear that much still needs to be discovered. The 'stem cell-like' cell lines being reported are still not satisfactorily used in regenerative medicine, due to reasons such as heterogeneity and chromosomal instability. As a result, investigators have searched for alternate source of cells that can be used for regenerative medicine. This approach has uncovered a range of adult stem cells and adult progenitor cells that have utility in both human and veterinary medicine. Here, we review a range of stem cells, from ESC to induced pluripotent stem cells, and discuss their potential application in the field of regenerative medicine.

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Section: Adult Stem Cellsmentioning

confidence: 99%

Stem Cells and Regenerative Medicine in Domestic and Companion Animals: A Multispecies Perspective

Gonçalves¹,

Ambrósio²,

Piedrahita

2014

Reprod Domestic Animals

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“…After digestion, the solution was filtered with a 70-mm cell strainer (BD Biosciences, San Jose, CA) to collect the NPCs in the filtrate, which were centrifuged and resuspended in a freezing medium [DMEM/F-12 GlutaMAX (Gibco, Carlsbad, CA) with 10% fetal calf serum (FCS; Gibco) and 10% dimethyl sulfoxide (DMSO; Merck, Darmstadt, Germany)] and cryopreserved in liquid nitrogen until further use. BMSCs were harvested from the diaphysis of humeri, according to a previously described protocol, 23 and expanded in aMEM + 10% FCS Gold (PAA Laboratories, Pasching, Austria) + 1% penicillin/streptomycin (pen/strep) until subconfluency, at which time BMSCs were trypsinized and resuspended in the freezing medium for cryopreservation in liquid nitrogen at P0.…”

Section: Methodsmentioning

confidence: 99%

Conditioned Medium Derived from Notochordal Cell-Rich Nucleus Pulposus Tissue Stimulates Matrix Production by Canine Nucleus Pulposus Cells and Bone Marrow-Derived Stromal Cells

Vries

Potier

Doeselaar

et al. 2015

Tissue Engineering Part A

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Objectives: Conditioned medium derived from notochordal cell-rich nucleus pulposus tissue (NCCM) was previously shown to have a stimulatory effect on bone marrow stromal cells (BMSCs) and nucleus pulposus cells (NPCs) individually, in mixed species in vitro cell models. The objective of the current study was to assess the stimulatory effect of NCCM on NPCs in a homologous canine in vitro model and to investigate whether combined stimulation with NCCM and addition of BMSCs provides a synergistic stimulatory effect. Methods: BMSCs and NPCs were harvested from chondrodystrophic dogs with confirmed early intervertebral disc (IVD) degeneration. NCCM was produced from NP tissue of nonchondrodystrophic dogs with healthy IVDs. BMSCs or NPCs alone (3 · 10 6 cells/mL) and NPCs + BMSCs (6 · 10 6 cells/mL; mixed 1:1) were cultured for 4 weeks in 1.2% alginate beads under base medium (BM), NCCM, or with addition of 10 ng/mL transforming growth factor-b1 (TGF-b1) as a positive control. Beads were assessed for glycosaminoglycan (GAG) and DNA contents by biochemical assays, GAG deposition by Alcian blue staining, and gene expression (aggrecan, versican, collagen 1 and 2, SOX9, A disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5), and matrix metalloproteinase 13 [MMP13]) with real-time quantitative RT-PCR. Results: NCCM increased NPC proliferation, proteoglycan production, and expression of genes associated with a healthy NP-like phenotype. BMSCs also showed increased proteoglycan production under NCCM, but these effects were not observed at the gene level. Combined stimulation of NPCs with NCCM and coculturing with BMSCs did not result in increased proteoglycan content compared to stimulation with NCCM alone. Discussion: NCCM stimulates matrix production by both NPCs and BMSCs and directs NPCs toward a healthier phenotype. NCCM is therefore promising for IVD regeneration and identification of the bioactive components will be helpful to further develop this approach. In the current study, no synergistic effect of adding BMSCs was observed.

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“…MSCs, NPCs and ACs were harvested from eight Beagle dogs (chondrodystrophic (CD1 through CD8; male, age 2.0 ± 0.3 years, weight 12.0 ± 1.3 kg (mean ± SD)). For each donor, bone marrow was collected and MSCs were isolated as described elsewhere [ 21 ]. When 80% confluence was reached (within 7 days), MSCs were cryopreserved at P0.…”

Section: Methodsmentioning

confidence: 99%

Effect of coculturing canine notochordal, nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration

Arkesteijn¹,

Smolders²,

Spillekom³

et al. 2015

Arthritis Res Ther

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IntroductionEarly degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cells (NPCs) decreases in a degenerating disc, mesenchymal stromal (stem) cells (MSCs) may be used for cell supplementation. In this study, using cells of one species, the regenerative potential of canine NCs was assessed in long-term three-dimensional coculture with canine NPCs or MSCs.MethodsCanine NCs and canine NPCs or MSCs were cocultured in alginate beads for 28 days under hypoxic and high-osmolarity conditions. Cell viability, cell morphology and DNA content, extracellular matrix production and expression of genes related to NC markers (Brachyury, KRT18) and NP matrix production (ACAN, COL2A1, COL1A1) were assessed after 1, 15 and 28 days of culture.ResultsNCs did not completely maintain their phenotype (morphology, matrix production, gene expression) during 28 days of culture. In cocultures of NPCs and NCs, both extracellular matrix content and anabolic gene expression remained unchanged compared with monoculture groups, whereas cocultures of MSCs and NCs showed increased glycosaminoglycan/DNA. However, the deposition of these proteoglycans was observed near the NCs and not the MSCs. Brachyury expression in the MSC and NC coculture group increased in time. The latter two findings indicate a trophic effect of MSCs on NCs rather than vice versa.ConclusionsNo regenerative potential of canine NCs on canine NPCs or MSCs was observed in this study. However, significant changes in NC phenotype in long-term culture may have resulted in a suboptimal regenerative potential of these NCs. In this respect, NC-conditioned medium may be better than coculture for future studies of the regenerative potential of NCs.Electronic supplementary materialThe online version of this article (doi:10.1186/s13075-015-0569-6) contains supplementary material, which is available to authorized users.

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Update on canine MSC markers

Cited by 8 publications

References 12 publications

Stem Cells and Regenerative Medicine in Domestic and Companion Animals: A Multispecies Perspective

Stem Cells and Regenerative Medicine in Domestic and Companion Animals: A Multispecies Perspective

Conditioned Medium Derived from Notochordal Cell-Rich Nucleus Pulposus Tissue Stimulates Matrix Production by Canine Nucleus Pulposus Cells and Bone Marrow-Derived Stromal Cells

Effect of coculturing canine notochordal, nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration

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