Xu R H, Rao H M, Zhu Y P
Basic Research Section, Shanghai Institute of Traumatology and Orthopedics, Shanghai Second Medical University.
Zhonghua Wai Ke Za Zhi. 1994 Mar;32(3):190-2.
Fibroblasts were harvested by in vitro culture of split-thickness skin of New-Zealand rabbit. Some opaque bone nodules were formed in the culture flasks when the fibroblasts underwent subculture for 8 days. Some of the nodules might enlarge, extend and form trabecula-like or flake-like structures, when the fibroblasts underwent subculture for 37 days. The above mentioned structures revealed golden-yellow fluorescence under the blue-violet light fluorescence microscope when the specimens were labeled with Tetracycline intravitally, denoting that they were newly formed bone tissues. In these structures, the calcium salt staining and collagen staining also showed positive reaction, fully reflecting the calcium and collagen composition. Osteogenesis caused by dermal fibroblast cultured in vitro was thus verified. There were evenly scattered oval spots in the osteogenic areas, after the specimens were treated with Alcina Blue-Sirius Red stain. Their diameter was about 10 microns, and they stained yellow color by Picric Acid. Therefore, these spots colored yellow with Picric Acid might be the dermal epithelia (and/or endothelia), which induced the fibroblasts to form the bone tissues.
采用新西兰兔断层皮肤进行体外培养收获成纤维细胞。当成纤维细胞传代培养8天时，培养瓶中形成一些不透明的骨结节。当成纤维细胞传代培养37天时，部分结节可能会增大、延伸并形成小梁状或片状结构。当标本用四环素进行体内标记后，上述结构在蓝紫光荧光显微镜下呈现金黄色荧光，表明它们是新形成的骨组织。在这些结构中，钙盐染色和胶原染色也呈阳性反应，充分反映了钙和胶原的组成。由此证实了体外培养的真皮成纤维细胞可诱导成骨。标本经阿尔辛蓝-天狼星红染色后，在成骨区域有均匀散在的椭圆形斑点。它们的直径约为10微米，经苦味酸染色呈黄色。因此，这些经苦味酸染成黄色的斑点可能是诱导成纤维细胞形成骨组织的真皮上皮（和/或内皮）。
