Characteristics of Primary Osteoblast Culture Derived from Rat Fetal Calvaria

大鼠胚胎頭顱骨取出之成骨細胞初代培養的特性之研究

何美泠(Mei-Ling Ho) ； 張瑞根(Je-Ken Chang) ； 莊麗月(Lea-Yea Chuang) ； 許勝光(Hseng-Kuang Hsu) ； 王國照(Gwo-Jaw Wang)

《The Kaohsiung Journal of Medical Sciences》 15卷5期 (1999/05) Pp. 248-255

https://doi.org/10.6452/KJMS.199905.0248

摘要

初代成骨細胞培養比成骨細胞株更能表現正常成骨細胞之表型特質，所以可以利用來做為體外探討骨細胞功能的實驗工具。本研究之初代成骨細胞是取自大白鼠胚胎頭顱骨之頂骨。將我們所培養之初代成骨細檢測各種不同的成骨細胞特性，同時亦測試纖維母細胞作為對照。我們分別在成骨細胞長滿後2週或3週作鹼性磷酸醴綉 (Alkaline phosphatase, ALP) 染色及von Kossa 染色來檢試ALP之表現及其後所發生之骨基質礦質化情形。結果顯示成細胞培養表現明顯的ALP及von Kossa染色，而纖維母細胞則為陰性反應，顯示本研究所建立的成骨細胞培養確能表現出典型的初代成骨細胞基因表型而與纖維母細胞不。我們更在不同劑量之前列腺素E2(PGE2)ketorolac之處理後測定初代成骨細胞及纖維母細胞之ALP活性的變化。結果顯示PGE2及ketorolac有劑量依賴性的刺激成骨細胞內ALP活性，而纖維母細胞則不管在對照組或各種劑量之藥物處理組均表現非常低之ALP活性。這些潔果顯示PGE2可能參與成骨細胞之份化作用，而且ketorolac對成骨細胞ALP性之刺激性作用可能非透過PGE2而達到作用的。這些成骨細胞對上述兩種藥約之反應也可作為由大鼠頭顱骨所取出之成骨細胞的特性。

並列摘要

Pirmary osteoblast cultures, which reflect more phenotypic properties of normal osteoblasts than osteoblastic cell lines, can be used as an experimental tool for investigating the osteoblastic functions in vitro. Primary osteoblast cultures wre obtained from the parietal bones of calvaria of fetal rats in this study. Differential characteristics of osteoblasts in our culture system were examined and fibroblast cultures were also tested for comparison. We tested the alkaline phosphatase (ALP) and von Kossa stains on osteoblast and fibroblast cultures ot examine the expression of ALP and the subsequent matrix mineralization occurred at 2 and 3 weeks after cell confluence respectively. The results showed that ostebloast cultures revealed obvious positive stains of ALP and von Kossa, while fibroblast cultures reveraled negative stains, suggesting the osteoblast culture system used in this study reflects the typical phenotypes of primary osteoblasts but not fibroblasts. We tested the ALP activities following various doses of PGE2 or ketorolac treatments in primary osteoblast and fibroblast cultures. The results showed that PGE2 and ketorolac stimulated intracellular ALP activities of osteoblasts in dose dependent fashions, while very low ALP activities were detected in that PGE2 may be involved in osteoblastic differentiation and the stimulatory effect of detorolac on osteoblastic ALP activity may not be PGE2 mediated. The responses of osteoblasts to both agents can be as the characteristics of primary osteoblast derived from rat calvaria.

並列關鍵字

osteoblast ； alkaline phosphatase ； mineralization