Comparative Study of Bone Cell Culture Methods for Tissue Engineering Applications

Abstract

Cells are used in bone tissue engineering applications to facilitate new bone formation in implants. Enzymatic digestion and marrow removal by either centrifugal force or syringe are three methods used to isolate the cells for culture, but each technique has benefits and drawbacks. This comparative study evaluated the effects of the three cell isolation techniques on the attachment, proliferation, and mineralization of rat bone cells. Cells were isolated, seeded, and cultured following standard protocols for each isolation method. Quantitative assays to determine metabolic activity, lactic acid production, glucose consumption, and amounts of intracellular protein, alkaline phosphatase activity, and extracellular calcium were performed. In addition, cell morphology and viability were examined qualitatively. The results indicate that the cell isolation method affects the attachment, proliferation, and type of tissue formed by cells cultured under identical conditions.

Effects of Tinospora cordifolia (Menispermaceae) on the proliferation, osteogenic differentiation and mineralization of osteoblast model systems in vitro

“…This part of the study which involves the usage of animals was carried out following ethical guidelines and is approved by the institutional animal ethics committee (IAEC approval No: BUB/ZOO/IAEC/ANIM/2009/16). The animals were sacrificed by cervical dislocation under ether anesthesia and osteoblasts isolated from femur by following the method of Bard et al (1972) and as modified by Thomas et al (2004) and Zhu et al (2010). The primary osteoblast cells were cultured in ready to use sterile liquid Dulbecco's minimum essential medium (DMEM AL007, Himedia, India) supplemented with 1X antibiotic antimycotic solution and 10% fetal bovine serum.…”

Selective Estrogen Receptor Modulator and prostimulatory effects of phytoestrogen β-ecdysone in Tinospora cordifolia on osteoblast cells

“…This part of the study which involved the usage of animals was carried out following ethical guidelines specified by Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Ministry of Environment and Forest and was approved by the Institutional Animal Ethics Committee (IAEC) (Approval No – BUB/IAEC/MB&BT/Dr.MSP/dated 09.05.11). The animals were sacrificed by CO 2 asphyxiation and the primary osteoblast cells were isolated from rat femur as described earlier [20] , [21] , [22] .…”

Material properties and bone marrow stromal cells response to In situ crosslinkable RGD‐functionlized lactide‐co‐glycolide scaffolds

“…Bone marrow stromal (BMS) cells were isolated from the bone marrow of young adult male Wistar rats 40, 41. After euthanasia, the femurs and tibias were aseptically excised from the hind limbs, cleaned of soft tissue, and washed in Dulbecco's Modified Eagle Medium (DMEM; 4.5 g/L glucose with L ‐glutamine and without sodium pyruvate; Cellgro; Herndon, VA) containing 5 times higher gentamicin sulfate (GS; Sigma; 100 μg/mL) concentration than for cell culture to avoid contamination during harvest.…”