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.

Material Properties and Cytocompatibility of Injectable MMP Degradable Poly(lactide ethylene oxide fumarate) Hydrogel as a Carrier for Marrow Stromal Cells

“…Bone Marrow Stromal Cell Isolation. BMS cells were isolated from the bone marrow of young adult male Wistar rats. , After euthanasia, the femurs and tibias were aseptically excised from the hind limbs, cleaned of soft tissue, and washed in DMEM 5 times higher than for cell culture concentration of gentamicin sulfate (GS; Sigma; 100 μg/mL) to avoid contamination during the harvest. Plugs of marrow were extracted by cutting the distal ends of tibias and the proximal ends of femurs to expose the marrow cavity.…”

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.…”

Hallazgo y caracterización de células "Stem" mesenquimales en núcleo pulposo de disco intervertebral cervical degenerado. Comparación con las células obtenidas de médula ósea de los mismo pacientes y resultados previos en la región lumbar

“…Aunque diferentes autores sugieren que las células obtenidas de uno y otra son idénticas, tanto en el potencial de diferenciación como de las características fenotípicas. Thomas et al [168] han llegado a la conclusión que las células aisladas por medio enzimático poseen mayor actividad metabólica, nivel de proteínas intracelulares y producción de calcio comparado con las aspiradas. los vicios de las CFU-F de la médula como un todo, excepto que estas células se obtienen con una eficacia menor que con los protocolos iniciales.…”