Observation and quantitative analysis of rat bone marrow stromal cells cultured in vitro on newly formed transparent β-tricalcium phosphate

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Abstract

To observe living cell morphology on ceramics by light microscopy, we fabricated a new material—transparent β - tricalcium phosphate (t-β TCP) ceramic—for the purpose of serving as a tissue culture substrate. Bone marrow stromal cells (BMSCs) were obtained from rat femora and cultured on both t-β TCP ceramic disks and culture grade polystyrene (PS) dishes in an osteogenic medium. After 1 day of culture, cell attachment and spreading on both the t-β TCP and PS substrata were equally and clearly detected by ordinary light microscopy. After 14 days of culture, extensive cell growth, alkaline phosphatase (ALP) staining, and bone mineral deposition could be detected on both substrata. In addition, quantitative biochemical analyses revealed high DNA content, ALP activity, and osteocalcin content of these cultures. This experiment is significant in that all of the results were similarly observed on both the t-β TCP and PS substrata, indicating the excellent properties of β TCP ceramics for BMSCs culture towards osteogenic differentiation.