Conditioned medium: a new alternative for cryopreservation of equine umbilical cord mesenchymal stem cells.

Cryopreservation is a feasible alternative to maintaining several cell lines, particularly for immediate therapeutic use, transportation of samples, and implementation of new in vitro studies. This work parts from the hypothesis that the medium of cryopreservation composed by 90% of conditioned medium (CM) supports cryopreservation of equine umbilical cord intervascular matrix mesenchymal stem cells (UCIM-MSCs), allowing the maintenance of the biological properties for the establishment of cell banks intended for therapeutic use and in vitro studies. Thus, we evaluated the viability, apoptosis/necrosis rates, immunophenotypic profile (IP), chromosomal stability, clonicity, and differentiation potential of UCIM-MSCs cryopreserved with four different mediums (with FBS: M1, M3, M4 and without FBS: M2). After 3 months of cryopreservation, samples were thawed and analyzed. The potential of differentiation in the mesodermal lineages, clonicity, and the chromosomal stability were maintained after cryopreservation of UCIM-MSCs with medium containing FBS. Changes (P < 0.05) at IP for some markers were observed at cells cryopreserved with medium M1-M3. Only the UCIM-MSCs cryopreserved with the CM (M4) had similar viability post-thaw (P = 0.23) when compared with fresh cells. We proved the hypothesis that the medium of cryopreservation containing CM supports the cryopreservation of UCIM-MSCs, at the experimental conditions, being the medium that better maintains the biological characteristics observed at fresh cells. Thus, future studies of UCIM-MSCs secretome should be conducted to better understand the beneficial and protective effects of the CM during the freezing process.

Bovine endometrial cells: a source of mesenchymal stem/progenitor cells.

Endometrial mesenchymal stem/progenitor cells (eMSCs) are multipotent cells known to modulate the immune system and have clinical application for human and animal health. This makes these bovines cells attractive for dual use as cellular therapy and experimental model. The aim of this study was to isolate, evaluate the differentiation potential, immunophenotypic and immunocytochemistry characteristics, chromosomal stability, cloning efficiency, and cryopreservation response of bovine eMSCs collected in two phases of the estrous cycle. For this, cells were isolated and submitted to differentiation for adipogenic and osteogenic lineage. The cells were then characterized by flow cytometer (FC) (vimentin, CD29, CD44, MHC-II, CD34) and immunocytochemistry (vimentin, pan-cytokeratin, CD44) and submitted to cytogenetic and cloning efficiency assay. The cells were also cryopreserved using two different medium of cryopreservation and analyzed by FC for viability, necrosis, late-apoptosis + necrosis, and initial apoptosis rates before and after cryopreservation. We obtained homogeneous cell populations which have fibroblastic morphology and adherence to plastic. These cells expressed high levels of markers CD29, CD44, and vimentin, low expression levels for CD34 and no MHC-II. The cells were chromosomally stable (2n = 60) with high cloning efficiency and no difference (P > 0.05) between medium of cryopreservation or phase was observed after thawing. We showed the presence and differentiation potential of bovine eMSCs, with chromosomal stability and great response to cryopreservation with both medium, which has implications for build biobanks or development of new therapeutic approaches to combat uterine diseases or to study.

Intramuscular Transplantation of Allogeneic Mesenchymal Stromal Cells Derived from Equine Umbilical Cord.

BACKGROUND AND OBJECTIVES: Mesenchymal stromal cells (MSCs) have great therapeutic potential, particularly in the process of tissue repair and immunomodulation through the secretion of biomolecules. Thus, the aim of this study was to evaluate the hypothesis that intramuscular transplantation of allogeneic MSCs obtained from equine umbilical cord (UC-MSCs) is safe, demonstrating that this is a suitable source of stem cells for therapeutic use. METHODS AND RESULTS: For this, UC-MSCs were cultured, characterized and cryopreserved for future transplantation in six healthy mares. On day 0, transplantation of three million UC-MSCs diluted in Hank's Balanced Solution (HBSS) was performed on right and left sides of the rump muscle. As a control, HBSS injections were performed caudally in the same muscle. Muscle biopsies were obtained as a control 30 days before transplantation (D-30). The biopsies were collected again on day 2 (left side) and day 7 (right side) post transplantation and examined histologically. All procedures were preceded by ultrasound examination and blood sampling. Hematologic evaluation remained within normal limits and no differences were observed between time points (p>0.05). Ultrasound examination was suggestive of inflammation 48 hours after transplantation in both groups (control and treated). At histological evaluation it was found only discrete inflammation signals between D-30×D2 (p<0.05) in the treated group, without differences (p> 0.05) between the groups at different time points. CONCLUSIONS: Equine UC-MSCs under the experimental conditions did not promote severe inflammation that causes tissue damage or lead to its rejection by the host organism and therefore has a good potential for clinical use.

Inércia uterina e maceração fetal após aplicação incorreta do acetato de medroxiprogesterona em gata: relato de caso / Uterine inertia and fetal maceration after misapplication of medroxyprogesterone in cat: case report / Inercia uterina y la maceración del feto después de la aplicación errónea de medroxiprogesterona in la gata: reporte de un caso

O acetato de medroxiprogesterona (MA) é administrado em cadelas e gatas para supressão do estro. Porém, seu uso na gestação pode levar à distocia, salientando a importância em discutir os seus prováveis mecanismos de ação. Avaliou-se o caso de uma gata com secreção vaginal purulenta e não desencadeamento do parto, após aplicação do MA na gestação. Foi diagnosticada mace ração fetal e realizada ovariosalpingohisterectomia (OSH), com resolução da afecção. Os mecanismos pelos quais os progestágenos promovem a quiescência uterina ainda não são completamente conhecidos. Em outras espécies sugere-se ação nos níveis intracelulares de cálcio e de relaxina; nos receptores para ocitocina e na síntese de prostaglandina. No caso relatado, acredita-se que o MA inibiu a contração uterina e a abertura cervical desencadeou a maceração fetal.

The medroxyprogesterone acetate (MA) is administered to dogs and cats for suppression of oestrus. However, its use during pregnancy can lead to dystocia, emphasizing the importance of discussing their possible mechanisms of action. It was evaluated the case of a cat with purulent vaginal discharge and not onset the birth, after application of MA during pregnancy. Was diagnosed fetal maceration and performed ovariosalpingohisterectomy (OSH), with resolution of the disease. The mechanisms by which the incorrect administration of MA leads to not trigger parturition in cats are not fully understood. In other species suggest action on the intracellular calcium levels and relaxin; the receptors for oxytocin and prostaglandin synthesis. In the reported case, it is believed that MA inhibited uterine contraction and cervical opening triggered fetal maceration.

El acetato de medroxiprogesterona (MA) se administra a los perros y gatos para la supresión del estro. Sin embargo, su uso durante el embarazo puede causar distocia, haciendo hincapié en la importancia de discutir sus posibles mecanismos de acción. Evaluado el caso de un gato con la secreción vaginal purulenta y no desencadenar el nacimiento, después de la aplicación del MA en el embarazo. Fue diagnosticado maceración fetal y ovariosalpingohisterectomy (OSH) celebrada el trabajo, con la resolución de la enfermedad. Los mecanismos por los que la progesterona promueve reposo uterino no están todavía completamente conocidos. En otras especies sugieren acción sobre los niveles de calcio intracelular y relaxina; los receptores de la oxitocina y la síntesis de prostaglandinas. En nuestro caso, se cree que MA inhibió la contracción uterina y la abertura cervical desencadena la maceración fetal.

Inércia uterina e maceração fetal após aplicação incorreta do acetato de medroxiprogesterona em gata: relato de caso / Uterine inertia and fetal maceration after misapplication of medroxyprogesterone in cat: case report / Inercia uterina y la maceración del feto después de la aplicación errónea de medroxiprogesterona in la gata: reporte de un caso

O acetato de medroxiprogesterona (MA) é administrado em cadelas e gatas para supressão do estro. Porém, seu uso na gestação pode levar à distocia, salientando a importância em discutir os seus prováveis mecanismos de ação. Avaliou-se o caso de uma gata com secreção vaginal purulenta e não desencadeamento do parto, após aplicação do MA na gestação. Foi diagnosticada mace ração fetal e realizada ovariosalpingohisterectomia (OSH), com resolução da afecção. Os mecanismos pelos quais os progestágenos promovem a quiescência uterina ainda não são completamente conhecidos. Em outras espécies sugere-se ação nos níveis intracelulares de cálcio e de relaxina; nos receptores para ocitocina e na síntese de prostaglandina. No caso relatado, acredita-se que o MA inibiu a contração uterina e a abertura cervical desencadeou a maceração fetal.(AU)

The medroxyprogesterone acetate (MA) is administered to dogs and cats for suppression of oestrus. However, its use during pregnancy can lead to dystocia, emphasizing the importance of discussing their possible mechanisms of action. It was evaluated the case of a cat with purulent vaginal discharge and not onset the birth, after application of MA during pregnancy. Was diagnosed fetal maceration and performed ovariosalpingohisterectomy (OSH), with resolution of the disease. The mechanisms by which the incorrect administration of MA leads to not trigger parturition in cats are not fully understood. In other species suggest action on the intracellular calcium levels and relaxin; the receptors for oxytocin and prostaglandin synthesis. In the reported case, it is believed that MA inhibited uterine contraction and cervical opening triggered fetal maceration.(AU)

El acetato de medroxiprogesterona (MA) se administra a los perros y gatos para la supresión del estro. Sin embargo, su uso durante el embarazo puede causar distocia, haciendo hincapié en la importancia de discutir sus posibles mecanismos de acción. Evaluado el caso de un gato con la secreción vaginal purulenta y no desencadenar el nacimiento, después de la aplicación del MA en el embarazo. Fue diagnosticado maceración fetal y ovariosalpingohisterectomy (OSH) celebrada el trabajo, con la resolución de la enfermedad. Los mecanismos por los que la progesterona promueve reposo uterino no están todavía completamente conocidos. En otras especies sugieren acción sobre los niveles de calcio intracelular y relaxina; los receptores de la oxitocina y la síntesis de prostaglandinas. En nuestro caso, se cree que MA inhibió la contracción uterina y la abertura cervical desencadena la maceración fetal.(AU)