Objectives: The aim of this study was to isolate feline dental pulp stem cells (fDPSCs) and characterize their clonogenic and proliferative abilities, as well as their multipotency, immunophenotype and cytogenetic stability. Methods: Dental pulp was isolated by explant culture from two cats <1 year old at post mortem. Their clonogenicity was characterized using a colony-forming unit fibroblast assay, and their proliferative ability was quantified with a doubling time assay in passages 2, 4 and 6 (P₂, P₄ and P₆, respectively). Multipotency was characterized with an in vitro trilineage differentiation assay in P₂, and cells were immunophenotyped in P₄ by flow cytometry. Chromosomic stability was evaluated by cytogenetic analysis in P₂, P₄ and P₆. Results: The fDPSCs displayed spindle and epithelial-like morphologies. Isolated cells showed a marked clonogenic capacity and doubling time was maintained from P₂ to P₆. Trilineage differentiation was obtained in one sample, while the other showed osteogenic and chondrogenic differentiation. Immunophenotypic analysis showed fDPSCs were CD45⁻, CD90⁺ and CD44⁺. Structural and numerical cytogenetic aberrations were observed in P₂–P₄. Conclusions and relevance: In this study, fDPSCs from two cats were isolated by explant culture and immunophenotyped. Cells displayed clonogenic and proliferative ability, and multipotency in vitro, and signs of chromosomic instability were observed. Although a larger study is needed to confirm these results, this is the first report of fDPSC isolation and in vitro characterization.

Platelet lysate (PL) is investigated as a potential replacement for fetal bovine serum (FBS) in cell culture. However, there is limited research on its impact on the immune profile of equine mesenchymal stromal cells (eMSCs). This study aimed to evaluate the effects of different PL formulations on the proliferative capacity, multipotentiality, and immune profile of equine adipose tissue-derived MSCs (eAD-MSCs). In vitro growth kinetics and trilineage differentiation of eAD-MSCs (n = 7) were assessed under three culture conditions: medium-concentration PL (MPL), high-concentration PL (HPL), and FBS as a control. The immune profile was evaluated by studying the expression of immunogenic receptors such as MHC I, MHC II, and immunomodulatory molecules IL-6, IL-10, and TNF-α, determined by gene expression, surface marker expression, and cytokine quantification. Both PL formulations, pooled from 5 donors, exhibited 3.3 and 6.5-fold higher platelet counts than baseline plasma for MPL and HPL, respectively. Higher concentrations of TGF-β and PDGF were found in both PL formulations compared to baseline. Furthermore, MPL and HPL subcultures demonstrated proliferative, clonogenic, and multipotent capacities similar to FBS. The immune profile of PL-cultured cells exhibited gene expression levels related to immunogenicity and immunomodulation similar to the reference condition, and the surface antigen presence of MHC II was also similar. However, HPL media exhibited higher IL-6, IL-10, and TNF-α concentrations in the culture supernatant. In conclusion, both PL media contained higher concentrations of growth factors compared to FBS, supporting the in vitro culture of eAD-MSCs with proliferative, clonogenic, and multipotent capacity similar to the reference medium. Nonetheless, PL usage led to a variation in the immunomodulatory cytokine microenvironment, with higher concentrations of IL-6, IL-10, and TNF-α in HPL media compared to MPL and FBS.

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.

Species of the genus Senecio have been traditionally employed with medical purposes and it has been demonstrated that some of them have anti-proliferative activity on cancer-derived cell lines. In South America S. madagascariensis is one of the most important representatives of the genus. Almost all species contain pyrrolizidine alkaloids (PA), substances that can lead to hepatic damage. Thus, the aim of this work was to determine the cytotoxic activity of a low PAs extract of S. madagascariensis on a cancer-derived cell line. A chloroform extract from plant material was produced and it was demonstrated to have a PAs concentration lower than 200 μg/g (w/w). Two stock solutions were made with DMSO and acetone as solvents and were diluted in culture medium (DMEM). Cells from urinary bladder transitional cell carcinoma T24 were cultured in 96 well plates (104 cells/well) and incubated for 24 hours with dilutions of the extracts at the following concentrations, 0.5 mg/mL, 0.25 mg/mL, 0.125 mg/mL and 0.06 mg/mL. Cell viability was evaluated by the MTT colorimetric technique. Both solutions of S. madagascariensis extracts (acetone and DMSO) showed cytotoxic activity. In both cases, the degree of cytotoxicity was dependent on the concentration of the solution.

La fibrina rica en plaquetas y leucocitos (L-PRF) es un producto sanguíneo de segunda generación que puede utilizarse como relleno biológico. Esto se debe a que la densa red de fibrina que se forma actúa como un andamio celular y proporciona factores de crecimiento de manera sostenida, lo que promueve la curación. En este estudio se evaluaron dos casos clínicos de pacientes que presentaron fístula oronasal (FON) atendidos en el Servicio de Odontoestomatología de la Facultad de Veterinaria, Udelar. Se extrajo el diente afectado, se cerró la FON mediante un colgajo mucoperióstico y se colocó L-PRF en la cavidad dental para promover la cicatrización. En los controles posoperatorios de los días 7, 14 y 30 se observaron resultados favorables, sin complicaciones ni recurrencias para ambos casos clínicos. El uso de L-PRF en la resolución quirúrgica de la FON resultó ser una técnica rápida, económica y sencilla, fácil de realizar en una clínica veterinaria, inofensiva y de bajo riesgo, resultó un novedoso tratamiento para el cierre de defectos alveolares.

Adipose tissue can be a source of bovine mesenchymal stromal/stem cells (MSCs). However, most studies do not incorporate post-mortem collection or in vitro differentiation of these cells from bovine fat and other sources into three mesodermal lineages. This study characterized and cryopreserved bovine adipose tissue-derived MSCs (AD-MSCs) collected post-mortem. Cells were isolated from cattle (Bos taurus) and cultured under standard conditions for ex vivo expansion and cryopreservation. Cell growth kinetics was determined by cell doubling time after consecutive cellular passages. Clonogenic capacity was determined using the colony-forming unit fibroblast (CFU-F) assay. The trilineage differentiation assay was performed to determine its in vitro multipotency. The in vitro proliferation and clonogenic capacity of bovine AD-MSCs was maintained across successive passages (P₂-P₆). Greater clonogenic capacity was found in fresh than post-cryopreservation cells. Cells showed multipotential capacity in adipose, cartilage and bone lineages. In conclusion, no changes were observed in the growth kinetics and colony-forming capacity of the successive passages evaluated. Clonogenic capacity was higher in fresh than post-cryopreserved cells.