Cardiac Specific Gene Expression Changes in Long Term Culture of Murine Mesenchymal Stem Cells

Murine MSCs are a readily available source of adult stem cells enabling extensive in vitro study of this cell population. MSCs have been described as multipotent, and have been proven capable of differentiation into several connective tissue types. Furthermore some studies have suggested an ability to differentiate into non-connective tissue cell types such as the cardiomyocyte. The aim of this study was to differentiate murine MSCs toward cardiac lineage with the commonly used method of culture with 5’ Azacytidine. Critically, baseline analysis of gene expression of passage four MSCs demonstrated expression of key cardiac markers including cardiac troponin T and I, and the ryanodine receptor. Furthermore, expression analysis of these genes changed with time in culture and passage number. However, there was no significant alteration when cells were subjected to a differentiation protocol. This study therefore highlights the importance of analyzing baseline cells extensively, and indicates the limitations in extrapolating data for comparison between species. Furthermore this data brings into question the efficacy of cardiac differentiation using MSCs

Feline mammary carcinoma stem cells are tumorigenic, radioresistant, chemoresistant and defective in activation of the ATM/p53 DNA damage pathway

AbstractCancer stem cells were identified in a feline mammary carcinoma cell line by demonstrating expression of CD133 and utilising the tumour sphere assay. A population of cells was identified that had an invasive, mesenchymal phenotype, expressed markers of pluripotency and enhanced tumour formation in the NOD-SCID mouse and chick embryo models. This population of feline mammary carcinoma stem cells was resistant to chemotherapy and radiation, possibly due to aberrant activation of the ATM/p53 DNA damage pathway. Epithelial–mesenchymal transition was a feature of the invasive phenotype. These data demonstrate that cancer stem cells are a feature of mammary cancer in cats

Relational quality and media use in interpersonal relationships

Author final draft doi:10.1177/1461444807080339This study examined the relationship between relational quality and media use in interpersonal relationships. In addition, the impacts of other potentially important variables such as sex and relationship type of participants and their partners were explored. College student participants focused on interaction experiences with an acquaintance, friend, romantic partner, or family member. Questions addressed the sex of relational partners, how much of participants’ total communication with relational partners is conducted in each of three media (i.e., face-to-face, phone, and internet), and the quality of relationships. Results indicated that participant sex and partner sex did not affect reported media use, whereas relationship type had significant effects on the extent to which face-to-face and telephone communication were used. Specifically, among the college students studied, face-to-face communication was used least with family members and the telephone was used most with family members. Relationships with acquaintances had the lowest relational quality and romantic relationships, while closer, were less satisfying than either family or friendship relationships. Same-sex relationships were perceived as more satisfying than cross-sex relationships. Finally, media use did not predict relational closeness or satisfaction. Results are discussed in light of previous research on mediated interpersonal communication and conceptualizations of the role of communication technology in one’s social life are highlighted

The activity of the feline thyroglobulin promoter is compromised by flanking adenoviral sequence

The use of human adenovirus (Ad) vectors for transcriptionally targeted thyroid gene therapy was investigated, aiming to develop a novel therapy for feline hyperthyroidism. Ad5 (El, E3 deleted) vectors were constructed in which the feline thyroglobulin promoter controls expression of the reporter gene chloramphenicol acetyl transferase (CAT). In FRTL-5 cells, CAT expression from adenoviral constructs harbouring the expression cassette was much reduced compared to controls transfected with the same cassette in a plasmid backbone, despite higher transduction efficiencies when viral vectors were used. Transfections with the “shuttle” plasmids utilised to create the adenoviral vectors also resulted in reduced expression compared to controls. In both viruses and shuttle plasmids, consistently lower expression was noted when the cassette was in the left to right orientation than the right to left. These results suggest cis acting elements in the flanking adenoviral sequences may compromise the activity of the feline thyroglobulin promoter and thus make Ad5 an unsuitable vector for transcriptionally targeted gene therapy in feline hyperthyroidism

Feline hyperthyroidism: advances towards novel molecular therapeutics

Feline hyperthyroidism is the most common endocrine disorder of the elderly cat. Traditionally, the disease is treated by surgical thyroidectomy, medical management with antithyroid drugs or radiation therapy using iodine-131. However, none of these treatments is ideal and molecular therapeutics may offer novel methods of treating the disease. This article reviews the background of, and preliminary investigations into, the development of a transcriptionally targeted somatic gene therapy strategy for the treatment of this feline condition

E. coli nitroreductase/CB1954: In vitro studies into a potential system for feline cancer gene therapy

Investigations were carried out to identify a suitable prodrug activating system for feline gene therapy with the eventual aim of treating feline thyroid disease and feline neoplasia. The E. coli nitroreductase (NTR)/CB1954 prodrug activating system was evaluated in vitro in feline cells by transient transfection with a nitroreductase expressing construct and subsequent treatment with the prodrug CB1954. The feline cells successfully expressed E. coli nitroreductase, which was able to activate the prodrug CB1954 resulting in cytotoxicity to both transformed and adjacent cells (a bystander effect) in vitro. In the absence of nitroreductase, CB1954 was non-toxic to feline cells. In addition, the nitroreductase gene was expressed in rat thyroid cells under the control of the cell type specific feline thyroglobulin promoter. This paper demonstrates that the E. coli nitroreductase/CB1954 system may be suitable for in vivo feline gene therapy, and further investigations are warranted

E. coli nitroreductase/CB1954: In vitro studies into a potential system for feline cancer gene therapy

Investigations were carried out to identify a suitable prodrug activating system for feline gene therapy with the eventual aim of treating feline thyroid disease and feline neoplasia. The E. coli nitroreductase (NTR)/CB1954 prodrug activating system was evaluated in vitro in feline cells by transient transfection with a nitroreductase expressing construct and subsequent treatment with the prodrug CB1954. The feline cells successfully expressed E. coli nitroreductase, which was able to activate the prodrug CB1954 resulting in cytotoxicity to both transformed and adjacent cells (a bystander effect) in vitro. In the absence of nitroreductase, CB1954 was non-toxic to feline cells. In addition, the nitroreductase gene was expressed in rat thyroid cells under the control of the cell type specific feline thyroglobulin promoter. This paper demonstrates that the E. coli nitroreductase/CB1954 system may be suitable for in vivo feline gene therapy, and further investigations are warranted

Characterisation and cardiac directed differentiation of canine adult cardiac stem cells

This study describes the isolation and characterisation of adult canine cardiac stem cells, and explores their ability to differentiate into cardiac myocytes. Direct comparisons are also made with available human data. Atrial cardiac explants were taken from dogs post-mortem and cultured to isolate adult stem cells. Cells were able to survive successive passages in serum-free media, were able to form cardiospheres, and under controlled culture conditions were capable of clonal expansion, demonstrating their ability for self-renewal. Characterisation of these cells demonstrated the following marker profile: c-kit, GATA 4 and flk-1 positive; cardiac troponin T and NKx2.5 low. Cardiac lineage directed differentiation was performed based on the published literature. Gene expression studies demonstrated that cardiac directed differentiation was partially achieved, with up-regulation of cardiac troponin T and NKx2.5, and down-regulation of c-kit and endothelial lineage markers. However the cells did not express the ryanodine receptor or β(1)-adrenergic receptors and did not contract spontaneously