Passage Number of 4T1 Cells Influences the Development of Tumour and the Progression of Metastasis in 4T1 Orthotopic Mice.

Background: The study was aimed to elucidate the influence of passage number of 4T1 cells for the development of the ideal tumour model. Methods: A total of 24 female BALB/c mice was divided equally into three groups: i) control (phosphate buffered saline [PBS] only); ii) group A (subjected to 4T1 cells of passage number 9) and iii) group B (subjected to 4T1 cells of passage number 10). The injections were introduced at the 3rd mammary pad of the mice. The net volume of the tumours was examined. Histopathological analysis was conducted to compare the extent of metastasis in the different groups of mice. Results: Group B had a higher net volume of 4T1 tumour as compared to group A (P = 0.042). The coefficient of variation in the net volume of 4T1 tumour for group A was higher (135.3%) as compared to group B (40.79%). Group A only exhibited metastasis on the lungs, liver and spleen whereas group B showed metastasis to the heart, spleen, lungs and liver. Conclusion: The use of 4T1 cells from passage number 10 is more ideal for the development of 4T1 tumour.

Analysis of serum and gene expression profile of cytokines (IL-6, TNF-α and TGF-ß1) in chronic hepatitis C virus infection.

Background: Chronic hepatitis C virus (HCV) infection is one of the major causes of liver cirrhosis and liver carcinoma. Studies have indicated that an imbalance of cytokine activities could contribute to the pathogenesis of chronic HCV infection. This study aimed to investigate serum levels and gene expression of cytokines (IL-6, TNF-α and TGF-ß1) in chronic HCV infection among Malay male subjects. Methods: Thirty-nine subjects were enrolled from various health clinics in Kelantan, Malaysia, and divided into two groups: patients with chronic HCV infection (HP) and healthy control (HS). The serum cytokines IL-6, TNF-a-were measured using Luminex assay, and serum TGF-ß1 was measured by ELISA. The mRNA gene expression for IL-6, TNF-α and TGF-ß1 was measured by real-time reverse transcriptase polymerase chain reaction (RT-PCR). Results: There were statistically significant differences in the mean serum levels of IL-6, and TGF-ß1 in HP compared to HS group (p = 0.0180 and p = 0.0005, respectively). There was no significant difference in the mean serum level of TNF-α in HP compared to HS group. The gene expression for the studied cytokines showed no significant differences in HP compared to HS group. Conclusion: Serum IL-6 was significantly associated with chronic HCV infection.

Extracellular Vesicles from Mesenchymal Stem Cells as Potential Treatments for Osteoarthritis.

Osteoarthritis (OA) is a chronic degenerative disorder of the joint and its prevalence and severity is increasing owing to ageing of the population. Osteoarthritis is characterized by the degradation of articular cartilage and remodeling of the underlying bone. There is little understanding of the cellular and molecular processes involved in pathophysiology of OA. Currently the treatment for OA is limited to painkillers and anti-inflammatory drugs, which only treat the symptoms. Some patients may also undergo surgical procedures to replace the damaged joints. Extracellular vesicles (EV) play an important role in intercellular communications and their concentration is elevated in the joints of OA patients, although their mechanism is unclear. Extracellular vesicles are naturally released by cells and they carry their origin cell information to be delivered to target cells. On the other hand, mesenchymal stem cells (MSCs) are highly proliferative and have a great potential in cartilage regeneration. In this review, we provide an overview of the current OA treatments and their limitations. We also discuss the role of EV in OA pathophysiology. Finally, we highlight the therapeutic potential of MSC-derived EV in OA and their challenges.

Isolation and enhancement of a homogenous in vitro human Hertwig's epithelial root sheath cell population.

Hertwig's epithelial root sheath (HERS) cells play a pivotal role during root formation of the tooth and are able to form cementum-like tissue. The aim of the present study was to establish a HERS cell line for molecular and biochemical studies using a selective digestion method. Selective digestion was performed by the application of trypsin-EDTA for 2 min, which led to the detachment of fibroblast-like-cells, with the rounded cells attached to the culture plate. The HERS cells displayed a typical cuboidal/squamous-shaped appearance. Characterization of the HERS cells using immunofluorescence staining and flow cytometry analysis showed that these cells expressed pan-cytokeratin, E-cadherin, and p63 as epithelial markers. Moreover, RT-PCR confirmed that these cells expressed epithelial-related genes, such as cytokeratin 14, E-cadherin, and ΔNp63. Additionally, HERS cells showed low expression of CD44 and CD105 with absence of CD34 and amelogenin expressions. In conclusion, HERS cells have been successfully isolated using a selective digestion method, thus enabling future studies on the roles of these cells in the formation of cementum-like tissue in vitro.