成骨细胞胞内胞外碱性磷酸酶含量比较

Comparison between the Extracellular and Intracellular Alkaline Phosphatase Level of the Primary Cultured Osteoblast Cells

目的通过比较原代培养的成骨细胞细胞内、外碱性磷酸酶含量,为提高碱性磷酸酶检测结果的准确性提供实验依据。方法取4只出生24 h内的大鼠胎鼠的颅骨,采用胰酶和Ⅰ型胶原酶交替消化的方法获得原代成骨细胞。分别取培养1 d、3d、5 d各5瓶细胞,直接取培养成骨细胞的培养液作为对照组,采用超声破碎的方法破碎细胞,收集细胞悬液,作为实验组,用碱性磷酸酶含量试剂盒检测收集到的2组液体。结果各时间点均是实验组数值高于对照组数值,并且差异有统计学意义(P<0.05)。结论原代成骨细胞细胞内的碱性磷酸酶含量,显著高于成骨细胞外的碱性磷酸酶含量,建议尽量采用将细胞破碎的方法进行ALP含量的检测,以提高结果的准确性。

Objective To compare the extracellular and intracellular alkaline phosphatase level of the primary cultured osteoblast cells, and to improve the accuracy of the experimental results. Methods Skulls from 4 fetal rats which were born within 24 hours were taken. Trypsin and collagenase I were used in turn to digest osteoblasts. 5 bottles of cells were taken at 1 d, 3 d and 5 d respectively. The culture solution was directly picked up as the control group. In the experimental group, osteoblasts were broken by ultrasonic sound and the cell suspension was collected. The two kind of liquid were tested by alkaline phosphatase detection kit. Results Alkaline phosphatase levels of the experimental group at each time point were significantly higher than those of the control group （P 〈 0.05 ）. Conclusion The intracellular alkaline phosphatase levels in the primary osteoblaststs are significantly higher than the extracellular level. In order to improve the accuracy of the resuits, we suggest using the disruption method to test the alkaline phosphatase level as possible as you can.