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Lummen, Belgium

Broeckx S.Y.,Global Stem Cell Technology | Maes S.,Valuepath | Maes S.,IDEXX Laboratories | Martinello T.,University of Padua | And 7 more authors.
Stem Cells and Development | Year: 2014

Besides the presence of somatic stem cells in hair follicles and dermis, the epidermis also contains a subpopulation of stem cells, reflecting its high regenerative capacity. However, only limited information concerning epidermis-derived epithelial-like stem/progenitor cells (EpSCs) is available to date. Nonetheless, this stem cell type could prove itself useful in skin reconstitution after injury. After harvesting from equine epidermis, the purified cells were characterized as EpSCs by means of positive expression for CD29, CD44, CD49f, CD90, Casein Kinase 2β, p63, and Ki67, low expression for cytokeratin (CK)14 and negative expression for CD105, CK18, Wide CK, and Pan CK. Furthermore, their self-renewal capacity was assessed in adhesion as well as in suspension. Moreover, the isolated cells were differentiated toward keratinocytes and adipocytes. To assess the regenerative capacities of EpSCs, six full-thickness skin wounds were made: three were treated with EpSCs and platelet-rich-plasma (EpSC/PRP-treated), while the remaining three were administered carrier fluid alone (PRP-treated). The dermis of EpSC/PRP-treated wounds was significantly thinner and exhibited more restricted granulation tissue than did the PRP-treated wounds. The EpSC/PRP-treated wounds also exhibited increases in EpSCs, vascularization, elastin content, and follicle-like structures. In addition, combining EpSCs with a PRP treatment enhanced tissue repair after clinical application. © Copyright 2014, Mary Ann Liebert, Inc. 2014. Source

Broeckx S.,Global Stem Cell Technology | Suls M.,Equine Veterinary Practice Dr Suls | Beerts C.,Global Stem Cell Technology | Vandenberghe A.,Equine Veterinary Practice Dr Suls | And 4 more authors.
Current Stem Cell Research and Therapy | Year: 2014

Cell-based therapies, such as treatments with mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP) are thought to have beneficial effects on the clinical outcome of orthopedic injuries, but very few animal studies with large sample size are published so far. Therefore, the aim of this study was to assess the safety and report the clinical outcome of allogenic, immature or chondrogenic induced MSCs in combination with PRP for the treatment of degenerative joint disease (DJD) in 165 horses. MSCs and PRP were isolated from a 6-year-old donor horse and transplanted either in their native state or after chondrogenic induction in combination with PRP into degenerated stifle (n=30), fetlock (n=58), pastern (n=34) and coffin (n=43) joints. Safety was assessed by means of clinical evaluation and the outcome was defined as failure to return to work (score 0), rehabilitation (score 1), return to work (score 2) and return to previous level (score 3), shortly (6 weeks) after treatment or at 18 weeks for the patients that returned for long-term follow-up (n=91). No adverse effects were noticed, except for three patients who showed a moderate flare reaction within one week after treatment of the fetlock joint without long-term effects (1.8% of 165 horses). Already after 6 weeks, 45% (native MSCs) and 60% (chondrogenic induced MSCs) of the treated patients returned to work (—> score 2+3) and the beneficial effects of the treatment further increased after 18 weeks (78% for native MSCs and 86% for chondrogenic induced MSCs). With the odds ratio of 1.47 for short-term and 1.24 for long-term, higher average scores (but statistically not significant) could be noticed using chondrogenic induced MSCs as compared to native MSCs. For all three lower limb joints a higher percentage of the treated patients returned to work after chondrogenic induced MSC treatment, whereas the opposite trend could be noticed for stifle joints. Nevertheless, more protracted follow-up data should confirm the sustainability of these joints. © 2014 Bentham Science Publishers. Source

Spaas J.,Research Center for Movement Control and Neuroplasticity | Helsen W.F.,Research Center for Movement Control and Neuroplasticity | Adriaenssens M.,Elscolab | Broeckx S.,Global Stem Cell Technology | And 2 more authors.
Veterinary Journal | Year: 2014

There is general agreement that horses have dichromatic colour vision with similar capabilities to human beings with red-green colour deficiencies. However, whether colour perception has an impact on equine jumping performance and how pronounced the colour stimulus might be for a horse is unknown. The present study investigated the relationship between the colour of the fences (blue or green) and the show jumping performance of 20 horses ridden by two riders using an indoor and outdoor set of green and blue fences.In the indoor arena, significantly more touches and faults were made on blue fences in comparison to green fences (median difference of 2.5 bars). When only touched bars were included, a significant median difference of one bar was found. Mares (n=4) demonstrated more faults and had a significantly greater difference in touches and faults between the two colours than male horses (n=16). Repeating the same experiment with eight horses in an outdoor grass arena revealed no significant differences between the two colours. In order to draw any definite conclusions, more research concerning the colour perception, influence of contrast with the arena surface and sex of horse is required. © 2014 Elsevier Ltd. Source

Broeckx S.,Global Stem Cell Technology | Zimmerman M.,Equine Veterinary Practice Dr Suls | Crocetti S.,ETH Zurich | Suls M.,Equine Veterinary Practice Dr Suls | And 11 more authors.
PLoS ONE | Year: 2014

Degenerative joint disease (DJD) is a major cause of reduced athletic function and retirement in equine performers. For this reason, regenerative therapies for DJD have gained increasing interest. Platelet-rich plasma (PRP) and mesenchymal stem cells (MSCs) were isolated from a 6-year-old donor horse. MSCs were either used in their native state or after chondrogenic induction. In an initial study, 20 horses with naturally occurring DJD in the fetlock joint were divided in 4 groups and injected with the following: 1) PRP; 2) MSCs; 3) MSCs and PRP; or 4) chondrogenic induced MSCs and PRP. The horses were then evaluated by means of a clinical scoring system after 6 weeks (T1), 12 weeks (T2), 6 months (T3) and 12 months (T4) post injection. In a second study, 30 horses with the same medical background were randomly assigned to one of the two combination therapies and evaluated at T1. The protein expression profile of native MSCs was found to be negative for major histocompatibility (MHC) II and p63, low in MHC I and positive for Ki67, collagen type II (Col II) and Vimentin. Chondrogenic induction resulted in increased mRNA expression of aggrecan, Col II and cartilage oligomeric matrix protein (COMP) as well as in increased protein expression of p63 and glycosaminoglycan, but in decreased protein expression of Ki67. The combined use of PRP and MSCs significantly improved the functionality and sustainability of damaged joints from 6 weeks until 12 months after treatment, compared to PRP treatment alone. The highest short-term clinical evolution scores were obtained with chondrogenic induced MSCs and PRP. This study reports successful in vitro chondrogenic induction of equine MSCs. In vivo application of (induced) MSCs together with PRP in horses suffering from DJD in the fetlock joint resulted in a significant clinical improvement until 12 months after treatment. © 2014 Broeckx et al. Source

Borena B.M.,Ghent University | Borena B.M.,Ambo University | Meyer E.,Ghent University | Chiers K.,Ghent University | And 5 more authors.
Cellular Physiology and Biochemistry | Year: 2014

Background: Mammal skin plays a pivotal role in several life preserving processes and extensive damage may therefore be life threatening. Physiological skin regeneration is achieved through ongoing somatic stem cell differentiation within the epidermis and the hair follicle. However, in severe pathological cases, such as burn wounds, chronic wounds, and ulcers, the endogenous repair mechanisms might be insuffcient. For this reason, exogenous purifcation and multiplication of epithelial-like stem/progenitor cells (EpSCs) might be useful in the treatment of these skin diseases. However, only few reports are available on the isolation, purifcation and characterization of EpSCs using suspension cultures. Methods: In the present study, skin was harvested from 6 mares and EpSCs were isolated and purifed. In addition to their characterization based on phenotypic and functional properties, sphere formation was assessed upon isolation, i.e. at passage 0 (P0), and at early (P4) and late (P10) passages using different culture conditions. Results: On average 0.53 ± 0.28% of these primary skin-derived cells showed the capacity to form spheres and hence possessed stem cell properties. Moreover, signifcantly more spheres were observed in EpSC medium versus differentiation medium, corroborating the EpSCs' privileged ability to survive in suspension. Furthermore, the number of cells per sphere signifcantly increased over time as well as with subsequent passaging. Upon immunophenotyping, the presumed EpSCs were found to co-express cytokeratin (CK) 14, Casein kinase 2 beta and Major Histocompatibility Complex (MHC) I and expressed no pan CK and wide CK. Only a few cells expressed MHC II. Their differentiation towards keratinocytes (at P4 and P10) was confrmed based on co-expression of CK 14, Casein kinase 2 beta, pan CK and wide CK. In one of six isolates, a non-EpSC cell type was noticed in adherent culture. Although morphological features and immunohistochemistry (IHC) confrmed a keratinocyte phenotype, this culture could be purifed by seeding the cells in suspension at ultralow clonal densities (1 and 10 cells/cm2), yet with a signifcantly lower sphere forming effciency in comparison to pure EpSCs (P = 0.0012). Conclusion: The present study demonstrated sphere formation as a valuable tool to purify EpSCs upon their isolation and assessed its effectiveness at different clonal seeding densities for eliminating a cellular contamination. Copyright © 2014 S. Karger AG, Basel. Source

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