The canine parvovirus type 2 (CPV-2) causes an acute disease in dogs. It has been found to induce cell cycle arrest and DNA damage leading to cellular lysis. In this paper, we evaluated the apoptotic potential of the "new CPV-2a" in MDCK cells and elucidated the mechanism of the induction of apoptosis. The exposure of MDCK cells to the virus was found to trigger apoptotic response. Apoptosis was confirmed by phosphatidylserine translocation, DNA fragmentation assays, and cell cycle analysis. Activation of caspases-3, -8, -9, and -12 and decrease in mitochondrial potential in CPV-2a-infected MDCK cells suggested that the CPV-2a-induced apoptosis is caspase dependent involving extrinsic, intrinsic, and endoplasmic reticulum pathways. Increase in p53 and Bax/Bcl2 ratio was also observed in CPV-2a-infected cells.
Early transcriptome profile of goat peripheral blood mononuclear cells (PBMCs) infected with peste des petits ruminant's vaccine virus (Sungri/96) revealed induction of antiviral response in an interferon independent manner. Research in Veterinary Science, 124, 166-177.
The use of viruses for treatment of cancer overcomes the bottlenecks of chemotherapy and radiotherapy. Several viruses and their proteins have been evaluated for oncolytic effect. The VP3 protein (apoptin) of chicken anemia virus is one such protein with an inherent ability to lyse cancer and transformed cells while leaving normal cells unharmed. In the present study, the apoptosis inducing potential of VP3 protein of CAV was evaluated in human cervical cancer cell line (HeLa). It was found that in VP3-induced apoptosis, caspase-dependent intrinsic pathway plays an important role with the cleavage of poly (ADP-ribose) polymerase (PARP) and there was no evidence of involvement of death receptor-mediated extrinsic pathway. The results of this study provide intuitive information and strengthen the candidacy of apoptin as a viral oncotherapeutic agent.
The conventional method of Bone Marrow Stromal Cell (BMSC) isolation from live subjects are complex due to involvement of lot of expert personnel and pre and post operational medication and cares. Moreover, the concerning ethical issues also pose lots of restrictions for isolation of BMSC's making stem cells research restricted to certain elite laboratories only. This study aims to compare between the regular aspiration (invasive) method and an alternative, straight forward and non-invasive method of BMSC harvest. The BMSCs were harvested by both invasive and non-invasive methods and cultured in MSC (Mesenchymal Stem Cell) medium. The cells were undergone visual assessment of growth dynamics as well as identification and characterization of MSC cells, based on microscopic examination. Both of these tested methods successfully yielded significant amount of BMSC that were found to be identical in morphology, growth dynamics and in vitro cultural properties. Unlike the invasive method that requires a live animal, the non-invasive method relies on post-slaughtered bone and therefore obviates the requirement of skill personnel and setup. Eventually, the used bone from already dead animal no longer becomes the issue of conflict with animal ethics and welfare. The ease in cell harvest and lack of ethical barrier would definitely make BMSC harvest convenient and therefore, anticipated to be well accepted in resource poor laboratories around the globe.
Mesenchymal stem cells (MSCs) are one of the rarest sub-populations of bone marrow resident cells having inherent ability to differentiate into mesenchyme tissues e.g. bone, cartilage and adipose tissues. The natural selfrenewal ability and potential for lineage specific differentiation have made these cells an excellent material for research and therapy in regenerative medicine. But, successful isolation and in vitro expansion of these cells still remain the pivotal steps for majority of stem cell based applications. Various techniques have been successfully used for isolation of MSCs from laboratory animals, but those are difficult to apply for domestic species. Hence, harvesting MSCs from most domestic animals remains a real challenge. Here we have demonstrated an easy, convenient, low cost method of MSCs isolation from slaughtered animals. As a proof of concept, MSCs were isolated from bone marrow of 3 different species, namely, sheep, pig and goat. These cells expressed multiple markers and also retained their self-renewal potential, exhibited by successful sub-culturing over 30 passages. Moreover, MSCs expressed many pluripotency factors e.g. OCT4, Nanog, c-Myc, KLF2 and KLF4. This indicated that the bone marrow derived MSCs were at very early stage of commitment and therefore, possibly retained high plasticity. Since these cells are available from slaughtered animals, this circumvents the bioethical issues associated with invasive method of MSC isolation from bone marrow. This invaluable and easily adoptable method for isolation of MSCs from large domestic animal would encourage isolation process in other animals and help in future cell based researches and therapies in the field of regenerative medicine.
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