兔胚成骨细胞的原代培养与鉴定

Primary culture and identification of the rabbit embryo osteoblasts

目的 针对目前成骨细胞培养常用的两种方法即酶消化法和组织块法 ,探讨组织块法培养成骨细胞的可行性及其应用价值。方法 取产前 1周的胎兔颅骨 ,采用组织块法 ,在含L 抗坏血酸 5 0mg/L、β 甘油磷酸钠 10mmol/L的DMEM成骨细胞条件培养液中培养 ,所得细胞用倒置显微镜观察、透射电镜观察、碱性磷酸酶 (ALP)染色和钙化结节VonKossa染色等进行鉴定。结果 颅骨分离的细胞在成骨细胞条件培养液中培养 ,经鉴定证实为成骨细胞。结论 组织块法是分离、培养成骨细胞的理想方法之一 。

Objective To investigate the possibility of culturing osteoblast with tissue mess method and its applicating value based on the two techniques commonly used for the primary culture of osteoblasts: enzyme digestion method and tissue mess method.?Methods The cranial bone which harvested from the one week fetuses before born was cultured in the osteoblasts conditioned cultured liquid of DMEM containing 50mg/L of L ascorbic acid and β glycerophosphate(10mmol/L) with tissue mess method, then the acquired cells were identified with inverted microscope and transmission electron microscope observation?alkaline phosphorase stain,Von Kossa stain of calcified nodule.?Results The cells obtained from cranial bone were cultured in the osteoblasts conditioned cultured liquid and were identified to be osteoblasts by certain identification.?Conclusion The tissue mess method is the ideal method to obtain and culture osteoblasts and can be selected as the ideal seeded cells for the tissue engineering bone. It can be used for the corresponding study.