Current Status on Canine Foetal Fluid and Adnexa Derived Mesenchymal Stem Cells (original) (raw)
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A Comparative Study of Canine Mesenchymal Stem Cells Isolated from Different Sources
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In this study, we provide comprehensive analyses of mesenchymal stem cells (MSCs) isolated from three types of canine tissues: bone marrow (BM-MSCs), adipose tissue (AT-MSCs) and amniotic tissue (AM-MSCs). We compare their morphology, phenotype, multilineage potential and proliferation activity. The BM-MSCs and AM-MSCs showed fibroblast-like shapes against the spindle shape of the AT-MSCs. All populations showed strong osteogenic and chondrogenic potential. However, we observed phenotypic differences. The BM-MSCs and AT-MSCs revealed high expression of CD29, CD44, CD90 and CD105 positivity compared to the AM-MSCs, which showed reduced expression of all the analysed CD markers. Similarly, the isolation yield and proliferation varied depending on the source. The highest isolation yield and proliferation were detected in the population of AT-MSCs, while the AM-MSCs showed a high yield of cells, but the lowest proliferation activity, in contrast to the BM-MSCs which had the lowest isola...
PLoS ONE, 2021
Despite the increasing demand of cellular therapies for dogs, little is known on the differences between adult and fetal adnexa canine mesenchymal stem cells (MSCs), and data on their metabolic features are lacking. The present study aimed at comparing the characteristics of canine adipose tissue (AT) and umbilical cord matrix (UC) MSCs. Moreover, for the first time in the dog, the cellular bioenergetics were investigated by evaluating the two main metabolic pathways (oxidative phosphorylation and glycolysis) of ATP production. Frozen-thawed samples were used for this study. No differences in mean cell proliferation were found (P>0.05). However, while AT-MSCs showed a progressive increase in doubling time over passages, UC-MSCs showed an initial post freezing-thawing latency. No differences in migration, spheroid formation ability, and differentiation potential were found (P>0.05). RT-PCR analysis confirmed the expression of CD90 and CD44, the lack of CD14 and weak expression ...
Iranian Journal of Veterinary Medicine, 2013
BACKGROUND: Stem cell therapy in small animal medicine is still in its infancy and few in vitro and in vivo research projects regarding animal Mesenchymal Stem Cells (MSCs) have been carried out. On the other hand, Cell tracking is the first step of the cell-based therapies and is essential to recognize cell fate post transplantation. OBJECTIVES: The aim of this study was to isolate, characterize, and transduce cBM-MSCs. METHODS: Canine Bone Marrow-derived Mesenchymal Stem Cells (cBM- MSCs) were isolated from bone marrow of dogs and characterized based on morphology, differentiation capacities, and surface marker expressions. For the first time, we labeled cBM-MSCs by GFP-encoding lentiviral vector to track them. RESULTS: cBM-MSCs were successfully isolated and pro- liferated. Morphologically, these cells were similar to other MSCs from other sources and species and were able to differentiate into osteocytes and adipocytes. cBM-MSCs expressed surface marker CD44 but were not able to...
Stem Cells and Development, 2010
The canine model provides a large animal system to evaluate many treatment modalities using stem cells (SCs). However, only bone marrow (BM) protocols have been widely used in dogs for preclinical approaches. BM donation consists of an invasive procedure and the number and differentiation potential of its mesenchymal stem cells (MSCs) decline with age. More recently, umbilical cord was introduced as an alternative source to BM since it is obtained from a sample that is routinely discarded. Here, we describe the isolation of MSCs from canine umbilical cord vein (cUCV). These cells can be obtained from every cord received and grow successfully in culture. Their multipotent plasticity was demonstrated by their capacity to differentiate in adipocytic, chondrocytic, and osteocytic lineages. Furthermore, our results open possibilities to use cUCV cells in preclinical trials for many well-characterized canine model conditions homologs to human diseases.
Molecular Reproduction and Development, 2014
Fetal adnexa are a non-controversial source of mesenchymal stem cells (MSCs) that have high plasticity, a high proliferation rate, and the ability to differentiate towards multiple lineages. MSC populations have been characterized for their stemness and differentiation capabilities; more recent work has focused on MSC selection and on establishing predictable elements to discriminate the cells with the most potential for regenerative medicine. In this study, we cytogenetically and molecularly characterized and followed the in vitro proliferation and differentiation potential of earlypassage canine amniotic membrane MSCs (AM-MSCs) and umbilical cord matrix MSCs (UCM-MSCs) isolated from fetuses at early (35À40 days) and late (45À55 days) gestational ages. We found that cells from both fetal gestational ages showed similar features. In all examined cell lines, the morphology of proliferating cells typically appeared fibroblast-like. Population doublings, passaged up to 10 times, increased significantly with passage number. In both cell types, cell viability and chromosomal number and structure were not affected by gestational age at early passages. Passage-3 AM-and UCM-MSCs from both gestational phases also expressed embryonic (POU5F1) and mesenchymal (CD29, CD44) stemness markers, whereas hematopoietic and histocompatibility markers were never found in any sample. Passage-3 cell populations of each cell type were also multipotential as they could differentiate into neurocytes and osteocytes, based on cell morphology, specific stains, and molecular analysis. These results indicated that MSCs retrieved from the UCM and AM in the early and late fetal phases of gestation could be used for canine regenerative medicine.
Canine and Equine Mesenchymal Stem Cells Grown in Serum Free Media Have Altered Immunophenotype
Stem Cell Reviews and Reports, 2015
Mesenchymal stem cell (MSC) therapy is being increasingly used to treat dogs and horses with naturally-occurring diseases. However these animals also serve as critical large animal models for ongoing translation of cell therapy products to the human market. MSC manufacture for clinical use mandates improvement in cell culture systems to meet demands for higher MSC numbers and removal of xeno-proteins (i.e. fetal bovine serum, FBS). While serum-free media (SFM) is commercially available, its affects on MSC phenotype and immunomodulatory functions are not fully known. The objective of this study was to determine if specific MSC culture conditions, MSC expansion in HYPERFlasks® or MSC expansion in a commercially available SFM, would alter MSC proliferation, phenotype or immunomodulatory properties in vitro. MSCs cultured in HYPERFlasks® were similar in phenotype, proliferative capacity and immunomodulatory functions to MSCs grown in standard flasks however MSC yield was markedly increased. HYPERFlasks® therefore provide a viable option to generate greater cell numbers in a streamlined manner. Canine and equine MSCs expanded in SFM displayed similar proliferation, surface phenotype and inhibitory effect on lymphocyte proliferation in vitro. However, MSCs cultured in the absence of FBS secreted significantly less PGE 2 , and were significantly less able to inhibit IFNγ secretion by activated T-cells. Immunomodulatory functions altered by expansion in SFM were species dependent. Unlike equine MSCs, in canine adipose-derived MSCs, the inhibition of lymphocyte proliferation was not principally modulated by PGE 2. The removal of FBS from both canine and equine MSC culture systems resulted in altered immunomodulatory properties in vitro and warrants further investigation prior to moving towards FBS-free culture conditions.
Mesenchymal Stem Cells: Generalities and Clinical Significance in Feline and Canine Medicine
Animals
Mesenchymal stem cells (MSCs) are multipotent cells: they can proliferate like undifferentiated cells and have the ability to differentiate into different types of cells. A considerable amount of research focuses on the potential therapeutic benefits of MSCs, such as cell therapy or tissue regeneration, and MSCs are considered powerful tools in veterinary regenerative medicine. They are the leading type of adult stem cells in clinical trials owing to their immunosuppressive, immunomodulatory, and anti-inflammatory properties, as well as their low teratogenic risk compared with pluripotent stem cells. The present review details the current understanding of the fundamental biology of MSCs. We focus on MSCs’ properties and their characteristics with the goal of providing an overview of therapeutic innovations based on MSCs in canines and felines.
Cell Transplantation, 2012
Clinical trials utilizing bone marrow-derived mesenchymal stem cell (BM-MSC) therapies show promise for treating a variety of pathologic conditions. Paramount to optimization of such cell-based therapies is a thorough understanding of MSC biology. Despite the tremendous potential that exists for the clinical use of canine BM-MSCs in veterinary medicine, as well as in preclinical studies for human medicine, relatively little information exists regarding basic biological properties of the cells. In this study, we compared the importance of donor characteristics (age and harvest site) and ex vivo expansion on canine BM-MSC frequency (CFU-f) and differentiation potential. Advancing age was found to have a negative effect on CFU-f as well as osteogenic potential. Site of harvest was also found to have significant effects on MSC properties. MSCs obtained from the humerus were found at the lowest frequency and were least osteogenic compared to those harvested from the tibia, femur, and ili...
Canine Amniotic Membrane Derived Mesenchymal Stem Cells- Potential Sources for Regenerative Medicine
“Agriculture for Life, Life for Agriculture” Conference Proceedings
Canine mesenchymal stem cells (MSCs) can be defined with self renew potential and specific differentiation capacity. Amiotic membrane represent an important source of MSCs, which can be harvested by minimally invasive methods. The aim of our study was to evaluate the growth characteristics of canine amniotic membrane derived mesenchymal stem cells. The placenta samples were collected after cesarean section from healthy mixed breed dogs. MSCs isolation was performed using enzymatic method. Isolated cells were cultured in propagation medium: Dulbecco’s Modified Eagle’s Medium/F12 (DMEM/F12, Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% antibiotic-antimycotic (Sigma-Aldrich). The medium was changed after 4 days. The cell doubling number, cell proliferation capacity, cell doubling time, daily duplication rate and clonogenic efficacy were evaluated. Our study demonstrate the self renew potential of canine amniotic membrane derived mesenchymal stem cells, and can rep...
Comparative analysis of canine mesenchymal stem cells and bone marrow-derived mononuclear cells
Veterinary World, 2021
Background and Aim: Mesenchymal stem cells (MSCs), which have multi-lineage differentiation potentials, are a promising source for regenerative medicine. However, the focus of study of MSCs is shifting from the characterization of the differentiation potential to their secretion potential for cell transplantation. Tissue regeneration and the attenuation of immune responses are thought to be affected by the secretion of multiple growth factors and cytokines by MSCs. However, the secretion potential of MSCs profiling remains incompletely characterized. In this study, we focused on the secretion ability related and protein mRNA expression of dog adipose tissue-derived MSCs (AT-MSC), bone marrow (BM)-derived MSCs, and BM-derived mononuclear cells (BM-MNC). Materials and Methods: Real-time polymerase chain reaction analyses revealed mRNA expression of nine growth factors and seven interleukins in these types of cells and three growth factors protein expression were determined using Enzyme-linked immunosorbent assay. Results: For the BM-MNC growth factors, the mRNA expression of transforming growth factor-β (TGF-β) was the highest. For the BM-derived MSC (BM-MSC) and AT-MSC growth factors, the mRNA expression of vascular endothelial growth factor (VEGF) was highest. BM-MSCs and AT-MSCs showed similar expression profiles. In contrast, BM-MNCs showed unique expression profiles for hepatocyte growth factor and epidermal growth factor. The three types of cells showed a similar expression of TGF-β. Conclusion: We conclude that expression of cytokine proteins and mRNAs suggests involvement in tissue repair and protection.