Optimization of the Static Human Osteoblast/Osteoclast Co-culture System
Journal Title: Iranian Journal of Medical Sciences - Year 2018, Vol 43, Issue 2
Abstract
Osteoblasts (OBs) and osteoclasts (OCs) are 2 major groups of bone cells. Their cell-to-cell interactions are important to ensure the continuity of the bone-remodeling process. Therefore, the present study was carried out to optimize an OB/OC co-culture system utilizing the human OB cell line hFOB 1.19 and OCs extracted from peripheral blood mononuclear cells (PBMNCs). It was a 2-step procedure, involving the optimization of the OB culture and the co-culture of the OBs with PBMNCs at an optimum ratio. Firstly, pre-OBs were cultured to 90% confluency and the time required for differentiation was determined. OB differentiation was determined using the van Gieson staining to detect the presence of collagen and Alizarin Red for calcium. Secondly, OBs and OCs were co-cultured at the ratios of 1 OC: 1 OB, 1 OC: 4 OBs, 2 OCs: 1 OB, and 1 OC: 2 OBs. Tartrate-resistant acid phosphatase (TRAP) staining was used to detect the differentiation of the OCs. The results showed that collagen was present on day 1, whereas calcium was detected as early as day 3. Based on the result of TRAP staining, 1 OC: 2 OBs was taken as the most appropriate ratio. No macrophage colony-stimulating factor and receptor activator of the nuclear factor-κB ligand were added because they were provided by the OBs. In conclusion, these optimization processes are vital as they ensure the exact time point and ratio of the OB/OC co-culture in order to produce a reliable and reproducible co-culture system.
Authors and Affiliations
James Jam Jolly, Kok-Yong Chin, Mohd Fozi Nur Farhana, Ekram Alias, Kien Hui Chua, Wan Nuraini Wan Hasan, Soelaiman Ima-Nirwana
Keywords
Related Articles
- EP ID EP277624
- DOI -
- Views 109
- Downloads 0
Pain Relief with Wet Cupping Therapy in Rats is Mediated by Heat Shock Protein 70 and ß-Endorphin
Background: Wet cupping therapy is a complementary therapy in pain management. The mechanism of this therapy, however, needs further elucidation. Cells injured by wet cupping therapy seem to stimulate the expression of h...
Frequency of Smear-Negative Tuberculosis in Northwest Iran
Background: Microscopic smear examination is the most common test in tuberculosis (TB) detection. It is, however, not strong enough to identify TB in the majority of afflicted individuals; thus, a significant number of T...
Effects of Mindful Parenting Training on Clinical Symptoms in Children with Attention Deficit Hyperactivity Disorder and Parenting Stress: Randomized Controlled Trial
Background: Children with attention deficit hyperactivity disorder (ADHD) are at risk of impairment in multiple domains. This study aimed to investigate the effectiveness of mindful parenting training in reducing clinica...
The Effects of Size and Type of Vocal Fold Polyp on Some Acoustic Voice Parameters
Background: Vocal abuse and misuse would result in vocal fold polyp. Certain features define the extent of vocal folds polyp effects on voice acoustic parameters. The present study aimed to define the effects of polyp si...
Primordial Odontogenic Tumor of Mandible; A Case with Proposed Diagnostic Criteria
Primordial odontogenic tumor (POT) has been included as a new entity in a recent WHO (2017) classification of odontogenic tumors under benign mixed mesenchymal and epithelial tumors.1 Although there is controversy regard...