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Animals
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Stem-Cell-Based Therapies in Veterinary Medicine—Current Status and Perspectives
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Stem-Cell-Based Therapies in Veterinary Medicine—Current Status and Perspectives

A special issue of Animals (ISSN 2076-2615). This special issue belongs to the section "Veterinary Clinical Studies".

Deadline for manuscript submissions: closed (15 May 2024) | Viewed by 16428

Special Issue Editors

Dr. Carlos Eduardo Ambrósio

E-Mail Website
Guest Editor
Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga, São Paulo 13635-900, Brazil
Interests: stem cells; animal models; acquired and genetic diseases; placenta
Special Issues, Collections and Topics in MDPI journals
Dr. Daniele Santos Martins

E-Mail Website
Guest Editor
Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga, São Paulo 13635-900, Brazil
Interests: biobanking; stem cell lineages; animal models
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

In the last three decades, abundant information and data from different stem cell lineages in domestic and wild animals have been gathered in the literature. However, new sources of tissue and different types of cells from different species must be strongly described in future articles. Biobanking and new protocols of stem cell lineages are encouraged, as is the their use in animal models of diseases, both acquired and genetics.

We are pleased to invite researchers from around the world to submit your manuscripts on stem-cell-based therapies sharing possible new therapies using stem cell lines from different species in preclinical trials. Strong data from stem cells animal collections must be published to open new avenues of alternatives therapies.

Cell line biobanking may help to produce information for future use in conservative perspectives or even to treat reproduction problems and may guarantee cell lines for cloning purposes.

Another important aspect is the search for alternative therapies that will help to improve the quality of life of animals and their owners.

We invite researchers to submit your articles to this Special Issue.

Prof. Dr. Carlos Eduardo Ambrósio
Dr. Daniele Santos Martins
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Animals is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2400 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • stem cells
  • lineages
  • animal models
  • wild animals
  • domestic animals
  • biobanking

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Published Papers (7 papers)

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Research

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12 pages, 1868 KiB  
Article
Morphologic, Proliferative, and Cytogenetic Changes during In Vitro Propagation of Cat Adipose Tissue-Derived Mesenchymal Stromal/Stem Cells
by Agustina Algorta, Rody Artigas, Analía Rial, Uruguaysito Benavides, Jacqueline Maisonnave and Kevin Yaneselli
Animals 2024, 14(16), 2408; https://doi.org/10.3390/ani14162408 - 20 Aug 2024
Viewed by 1062
Abstract
Stem cell therapy in cat patients needs a high quantity of mesenchymal stromal/stem cells (MSCs) requiring in vitro propagation under culture conditions which may potentially impact cellular characteristics and genetic stability. This study aimed to assess the in vitro characteristics and cytogenetic stability [...] Read more.
Stem cell therapy in cat patients needs a high quantity of mesenchymal stromal/stem cells (MSCs) requiring in vitro propagation under culture conditions which may potentially impact cellular characteristics and genetic stability. This study aimed to assess the in vitro characteristics and cytogenetic stability of cat adipose tissue-derived MSCs (cAT-MSCs). For this purpose, morphological features, clonogenic potential, and proliferative capacity of cAT-MSCs were assessed at passages 2 (P2), P4, and P6. Multipotency and immunophenotype were evaluated. Cytogenetic analyses were conducted up to P6. The cAT-MSCs exhibited a spindle-shaped morphology in early passages. The doubling time increased from 2.5 days at P2 to 9.4 at P4 and 10.5 at P6, accompanied by the observation of nuclear abnormalities such as cluster formation, karyorrhexis, karyolysis, and a decline in the mitotic index at P4. Cells demonstrated multipotency capacity and were CD45−, CD90+, and CD44+. Metaphase analysis at P2 and P4 revealed some indications of structural instability such as gaps, breaks, deletions, duplications, and early chromatid segregation, but these alterations did not show an increase across passages. In conclusion, cAT-MSCs decreased their proliferative capacity after P4, accompanied by morphological alterations and signs of structural instability. Full article
(This article belongs to the Special Issue Stem-Cell-Based Therapies in Veterinary Medicine—Current Status and Perspectives)
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22 pages, 5897 KiB  
Article
Optimizing the Adipogenic Induction Protocol Using Rosiglitazone Improves the Physiological Parameters and Differentiation Capacity of Adipose Tissue-Derived Mesenchymal Stem Cells for Horses, Sheep, Dogs, Murines, and Humans
by Manuela Heimann, Mohamed I. Elashry, Michele C. Klymiuk, Asmaa Eldaey, Sabine Wenisch and Stefan Arnhold
Animals 2023, 13(20), 3224; https://doi.org/10.3390/ani13203224 - 15 Oct 2023
Cited by 1 | Viewed by 1978
Abstract
The investigation of adipose tissue-derived mesenchymal stem cells (ASCs) has received considerable interest in regenerative medicine. A nontoxic adipogenic induction protocol valid for cells of different mammalian species has not been described. This study aims to establish an adipogenic differentiation protocol suitable for [...] Read more.
The investigation of adipose tissue-derived mesenchymal stem cells (ASCs) has received considerable interest in regenerative medicine. A nontoxic adipogenic induction protocol valid for cells of different mammalian species has not been described. This study aims to establish an adipogenic differentiation protocol suitable for horses, sheep, dogs, murines, and human cells. An optimized rosiglitazone protocol, consisting of 5% fetal calf serum in Dulbecco’s Modified Eagle’s Medium, 10 μg/mL insulin, 0.55 μg/mL transferrin, 6.8 ng sodium selenite, 1 μM dexamethasone, and 1–5 μM of rosiglitazone, is compared to the 3-isobutyl-1-methylxantine (IBMX) protocol, where rosiglitazone was replaced with 0.5 mM IBMX and 0.2 mM indomethacin. Cell viability, cytotoxicity, a morphometric analysis of the lipid, and the expression of adipogenic markers for 14 days were assessed. The data revealed that using 5 µM of rosiglitazone promotes the adipogenic differentiation capacity in horse, sheep, and dog cells compared to IBMX induction. Meanwhile, marked reductions in the cell viability and cell number with the IBMX protocol were detected, and rosiglitazone increased the cell number and lipid droplet size, prevented apoptosis, and upregulated FABP-4 and Leptin expression in the cells of most of the species. Our data revealed that the rosiglitazone protocol improves the adipogenesis of ASCs, together with having less toxicity, and should be considered for cell reproducibility and clinical applications targeting obesity. Full article
(This article belongs to the Special Issue Stem-Cell-Based Therapies in Veterinary Medicine—Current Status and Perspectives)
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32 pages, 5287 KiB  
Article
Two Amnion-Derived Mesenchymal Stem-Cells Injections to Osteoarthritic Elbows in Dogs—Pilot Study
by Michal Domaniza, Marian Hluchy, Dasa Cizkova, Filip Humenik, Lucia Slovinska, Nikola Hudakova, Lubica Hornakova, Juraj Vozar and Alexandra Trbolova
Animals 2023, 13(13), 2195; https://doi.org/10.3390/ani13132195 - 4 Jul 2023
Viewed by 2411
Abstract
The aim of the study was to investigate the potential of cell-based regenerative therapy for elbow joints affected by osteoarthritis. Interest was focused on two intra-articular applications of amnion-derived mesenchymal stem cells (A-MSCs) to a group of different breeds of dogs with elbow [...] Read more.
The aim of the study was to investigate the potential of cell-based regenerative therapy for elbow joints affected by osteoarthritis. Interest was focused on two intra-articular applications of amnion-derived mesenchymal stem cells (A-MSCs) to a group of different breeds of dogs with elbow osteoarthritis (13 joints). Two injections were performed 14 days apart. We evaluated synovial fluid biomarkers, such as IFN-γ, IL-6, IL-15, IL-10, MCP-1, TNF-α, and GM-CSF, by multiplex fluorescent micro-bead immunoassay in the treated group of elbows (n = 13) (day 0, day 14, and day 28) and in the control group of elbows (n = 9). Kinematic gait analysis determined the joint range of motion (ROM) before and after each A-MSCs application. Kinematic gait analysis was performed on day 0, day 14, and day 28. Kinematic gait analysis pointed out improvement in the average range of motion of elbow joints from day 0 (38.45 ± 5.74°), day 14 (41.7 ± 6.04°), and day 28 (44.78 ± 4.69°) with statistical significance (p < 0.05) in nine elbows. Correlation analyses proved statistical significance (p < 0.05) in associations between ROM (day 0, day 14, and day 28) and IFN-γ, IL-6, IL-15, MCP-1, TNF-α, and GM-CSF concentrations (day 0, day 14, and day 28). IFN-γ, IL-6, IL-15, MCP-1, GM-CSF, and TNF- α showed negative correlation with ROM at day 0, day 14, and day 28, while IL-10 demonstrated positive correlation with ROM. As a consequence of A-MSC application to the elbow joint, we detected a statistically significant (p < 0.05) decrease in concentration levels between day 0 and day 28 for IFN-γ, IL-6, and TNF-α and statistically significant increase for IL-10. Statistical significance (p < 0.05) was detected in TNF-α, IFN-γ, and GM-CSF concentrations between day 14 and the control group as well as at day 28 and the control group. IL-6 concentrations showed statistical significance (p < 0.05) between day 14 and the control group. Full article
(This article belongs to the Special Issue Stem-Cell-Based Therapies in Veterinary Medicine—Current Status and Perspectives)
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17 pages, 15427 KiB  
Article
Initial Characterization of 3D Culture of Yolk Sac Tissue
by Vitória Mattos Pereira, Priscila Avelino Ferreira Pinto, Lina Castelo Branco Motta, Matheus F. Almeida, André Furugen Cesar de Andrade, Ana Paula Pinoti Pavaneli and Carlos Eduardo Ambrósio
Animals 2023, 13(9), 1435; https://doi.org/10.3390/ani13091435 - 22 Apr 2023
Cited by 2 | Viewed by 2740
Abstract
The role of the yolk sac (YS) in miscarriage is not yet clear, largely due to ethical reasons that make in vivo studies difficult to conduct. However, 3D cultures could provide a solution to this problem by enabling cells to be arranged in [...] Read more.
The role of the yolk sac (YS) in miscarriage is not yet clear, largely due to ethical reasons that make in vivo studies difficult to conduct. However, 3D cultures could provide a solution to this problem by enabling cells to be arranged in a way that more closely mimics the structure of the YS as it exists in vivo. In this study, three domestic species (porcine, canine, and bovine) were chosen as models to standardize 3D culture techniques for the YS. Two techniques of 3D culture were chosen: the Matrigel® and Hanging-Drop techniques, and the 2D culture technique was used as a standardized method. The formed structures were initially characterized using scanning electron microscopy (SEM), immunohistochemistry (IHC), and quantitative real-time PCR (RT-qPCR). In general, the 3D culture samples showed better organization of the YS cells compared to 2D cultures. The formed structures from both 3D methods assemble the mesothelial layer of YS tissue. Regarding the IHC assay, all in vitro models were able to express zinc and cholesterol transport markers, although only 3D culture techniques were able to generate structures with different markers pattern, indicating a cell differentiation process when compared to 2D cultures. Regarding mRNA expression, the 3D models had a greater gene expression pattern on the Hemoglobin subunit zeta-like (HBZ) gene related to the YS tissue, although no significant expression was found in Alpha-fetoprotein (AFP), indicating a lack of endodermal differentiation in our 3D model. With the initial technique and characterization established, the next step is to maintain the cultures and characterize the diversity of cell populations, stemness, functions, and genetic stability of each 3D in vitro model. Full article
(This article belongs to the Special Issue Stem-Cell-Based Therapies in Veterinary Medicine—Current Status and Perspectives)
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28 pages, 5393 KiB  
Article
Allogenic Synovia-Derived Mesenchymal Stem Cells for Treatment of Equine Tendinopathies and Desmopathies—Proof of Concept
by Inês Leal Reis, Bruna Lopes, Patrícia Sousa, Ana Catarina Sousa, Mariana Branquinho, Ana Rita Caseiro, Sílvia Santos Pedrosa, Alexandra Rêma, Cláudia Oliveira, Beatriz Porto, Luís Atayde, Irina Amorim, Rui Alvites, Jorge Miguel Santos and Ana Colette Maurício
Animals 2023, 13(8), 1312; https://doi.org/10.3390/ani13081312 - 11 Apr 2023
Cited by 5 | Viewed by 2718
Abstract
Tendon and ligament injuries are frequent in sport horses and humans, and such injuries represent a significant therapeutic challenge. Tissue regeneration and function recovery are the paramount goals of tendon and ligament lesion management. Nowadays, several regenerative treatments are being developed, based on [...] Read more.
Tendon and ligament injuries are frequent in sport horses and humans, and such injuries represent a significant therapeutic challenge. Tissue regeneration and function recovery are the paramount goals of tendon and ligament lesion management. Nowadays, several regenerative treatments are being developed, based on the use of stem cell and stem cell-based therapies. In the present study, the preparation of equine synovial membrane mesenchymal stem cells (eSM-MSCs) is described for clinical use, collection, transport, isolation, differentiation, characterization, and application. These cells are fibroblast-like and grow in clusters. They retain osteogenic, chondrogenic, and adipogenic differentiation potential. We present 16 clinical cases of tendonitis and desmitis, treated with allogenic eSM-MSCs and autologous serum, and we also include their evaluation, treatment, and follow-up. The concerns associated with the use of autologous serum as a vehicle are related to a reduced immunogenic response after the administration of this therapeutic combination, as well as the pro-regenerative effects from the growth factors and immunoglobulins that are part of its constitution. Most of the cases (14/16) healed in 30 days and presented good outcomes. Treatment of tendon and ligament lesions with a mixture of eSM-MSCs and autologous serum appears to be a promising clinical option for this category of lesions in equine patients. Full article
(This article belongs to the Special Issue Stem-Cell-Based Therapies in Veterinary Medicine—Current Status and Perspectives)
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Review

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16 pages, 312 KiB  
Review
Stem Cell Therapy for Aging Related Diseases and Joint Diseases in Companion Animals
by Yanmin Wang, Michael Alexander, Todd Scott, Desiree C. T. Cox, Augusta Wellington, Mike K. S. Chan, Michelle B. F. Wong, Orn Adalsteinsson and Jonathan R. T. Lakey
Animals 2023, 13(15), 2457; https://doi.org/10.3390/ani13152457 - 29 Jul 2023
Viewed by 3323
Abstract
Stem cell therapy is an attractive treatment for diseases in companion animals that cannot be treated by conventional veterinary medicine practices. The unique properties of stem cells, particularly the ability to differentiate into specific cell types, makes them a focal point in regenerative [...] Read more.
Stem cell therapy is an attractive treatment for diseases in companion animals that cannot be treated by conventional veterinary medicine practices. The unique properties of stem cells, particularly the ability to differentiate into specific cell types, makes them a focal point in regenerative medicine treatments. Stem cell transplantation, especially using mesenchymal stem cells, has been proposed as a means to treat a wide range of injuries and ailments, resulting in tissue regeneration or repair. This review aims to summarize the veterinary use of stem cells for treating age-related and joint diseases, which are common conditions in pets. While additional research is necessary and certain limitations exist, the potential of stem cell therapy for companion animals is immense. Full article
(This article belongs to the Special Issue Stem-Cell-Based Therapies in Veterinary Medicine—Current Status and Perspectives)

Other

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8 pages, 1992 KiB  
Case Report
Equine Corneal Wound Healing Using Mesenchymal Stem Cell Secretome: Case Report
by Alejandro Casado-Santos, Elsa González-Cubero, Maria Luisa González-Fernández, Yaiza González-Rodríguez, Mª Belén García-Rodríguez and Vega Villar-Suárez
Animals 2024, 14(13), 1842; https://doi.org/10.3390/ani14131842 - 21 Jun 2024
Viewed by 994
Abstract
Corneal ulcers are a common and potentially vision-threatening condition in horses that can be challenging to treat with conventional therapies alone. This case report describes the successful treatment of a non-healing corneal ulcer in a 28-year-old Hispano-Bretón mare using the secretome derived from [...] Read more.
Corneal ulcers are a common and potentially vision-threatening condition in horses that can be challenging to treat with conventional therapies alone. This case report describes the successful treatment of a non-healing corneal ulcer in a 28-year-old Hispano-Bretón mare using the secretome derived from adipose tissue-derived mesenchymal stem cells (ASCs). Despite initial treatment with antibiotics, anti-inflammatory drugs, and surgical debridement, the corneal ulcer failed to heal properly, exhibiting persistent epithelial defects and stromal complications. As an alternative regenerative approach, the ASC secretome, a rich source of trophic factors, cytokines, and extracellular vesicles, was topically administered to the affected eye. Remarkably, within one week of secretome treatment, the clinical signs of blepharospasm and epiphora resolved, and the corneal ulcer exhibited complete re-epithelialization, regained transparency, and reduced neovascularization. No recurrence was observed during the 1.5-year follow-up period. This case highlights the potential therapeutic benefits of the ASC secretome in promoting corneal wound healing and suggests its promise as a novel cell-free therapy for treating refractory corneal ulcers in horses. Full article
(This article belongs to the Special Issue Stem-Cell-Based Therapies in Veterinary Medicine—Current Status and Perspectives)
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