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.
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