Mesenchymal stem cells‐conditioned media (MSCs‐CM) contains several growth factors and cytokines, thus may be used as a better alternative to stem cell therapy, which needs to be elucidated. The present study was conducted to evaluate the therapeutic potential of caprine, canine, and guinea pig bone marrow‐derived MSCs‐CM in excision wound healing in a guinea pig model. MSCs were obtained from bone marrow, expanded ex vivo and characterized as per ISCT criteria. CM was collected assayed by western blot to ascertain the presence of important secretory biomolecules. Quantitative estimation by enzyme‐linked immunosorbent assay was done for a vascular epidermal growth factor (VEGF) and interleukin‐6 (IL‐6) in caprine MSCs‐CM and optimum time for collection of CM was decided as 72 hr. CM from all the species was lyophilized by freeze‐drying method. Full‐thickness (2 × 2 cm2) excision skin wounds were created in guinea pigs (six animals in each group) and respective lyophilized CM mixed with laminin gel was applied topically at weekly interval. On Day 28, histopathological examinations of healed skin were done by hemotoxylin and eosin staining. MSCs were found to secrete important growth factors and cytokines (i.e., VEGF, transforming growth factor‐β1, fibroblast growth factor‐2, insulin‐like growth factor‐1, stem cell factor, and IL‐6) as demonstrated by immunohistochemistry and western blot assay. It was found that allogenic and xenogenic application of CM significantly improved quality wound healing with minimal scar formation. Thus, MSCs‐CM can be used allogenically as well as xenogenically for quality wound healing.
The present study was designed to investigate the effect of MSCs-conditioned media (CM) on quality buffalo embryo production in vitro. MSCs were harvested from Wharton's jelly of 2-3 month old fetus and MSCs CM was collected. Immunocytochemistry and western blot assay revealed that MSCs secrete several important growth factors viz. FGF-2, IGF-1, LIF, TGF-β, and VEGF. Slaughterhouse derived culture grade cumulus oocyte complexes (COCs) were matured and fertilized in vitro. Presumptive zygotes were divided in four groups and cultured in vitro in respective media viz. group I (100% mSOF), Group II (100% Knockout Media DMEM+SR), Group III (50% CM + 50% mSOF), and group IV (100% CM). It was found that though the cleavage rate did not changed significantly (p < 0.05), but blastocyst rate was increased significantly (p < 0.05) in Group III and IV (24.24 ± 1.34 and 23.29 ± 1.25, respectively) compared to group I and II (16.04 ± 1.46 and 17.72 ± 0.94, respectively). Similarly, TCN was significantly (p < 0.05) higher in 50% CM and 100% CM replacement group (93.33 ± 1.91 and 92.13 ± 1.04, respectively) than the other two groups. It can be concluded from the study that MSCs secrete several important growth factors and MSCs-CM can be effectively used for enhancement of quality buffalo embryo production in vitro.
In most of domestic animals, semen biology is well studied, but it is not so well studied in camelids particularly in dromedary camels. In camelids, the ejaculated semen is highly viscous, so before its evaluation and processing for preservation, it is necessary to be liquefied. Our study was designed to evaluate the effect of three different enzymes (0.1% Collagenase, 0.5 X Accutase, 0.1% Trypsin-EDTA - Ethylene-diamine-tetra-acetic acid) on seminal viscosity at different time intervals along with its effects on seminal parameters. Semen was collected from the six adult healthy male camels using female camel as dummy. Bovine artificial vagina was used for the semen collection and ejaculate was kept at 37ºC for further processing after its collection. All three enzymes were diluted in 1x phosphate buffered saline (PBS) and the semen samples were kept at 37ºC. The samples were evaluated at different time intervals 0, 5, 10, 15, 30, 60, 120 and 240 min (T0–T240) for viscosity and it was evaluated by pipette method. Immediately after collection, the viscosity was very high, and no sperm mass motility was seen. However, after enzyme treatment viscosity was showed declining trend in all treatment groups compared to control. In trypsin treated group maximum motility was reached in 15 minutes. Live-dead percent was observed to be similar among treatment groups. Observations on liquefaction of semen revealed that use of enzymes decreasing the viscosity but with different rates.
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