Culture of rat primary osteoblasts using enzymatic digestion combined with tissue explant method

doi:10.3969/j.issn.2095-4344.2017.12.005

Abstract

Abstract:

BACKGROUND: Osteoblasts with high purity and activity are essential for bone metabolism research.
OBJECTIVE: To explore a simple and effective culturing method of primary osteoblasts.
METHODS: Osteoblasts were isolated from the parietal and frontal bones of newborn Sprague-Dawley rats using trypsin and collagenase digestion and tissue explant method. The morphology of osteoblasts was observed by inverted phase contrast microscope and transmission electron microscope; the cells was counted to draw the growth curve; the osteoblasts were identified by alkaline phosphatase BCIP/NBT staining and alizarin red staining.
RESULTS AND CONCLUSION: The cells showed spindle, triangle or polygon shapes, having two or three protrusions. There were abundant mitochondria and endoplasmic reticulum under electron microscope, which presented the typical characteristics of osteoblasts. The cell growth was slow intially, accelerating at the 3rd day,
and peaking at the 7th day. The cells were highly positive for alkaline phosphatase staining and were stained orangered through the alizarin red staining. To conclude, the cells isolated using enzymatic digestion combined with tissue explant method exhibit the typical characteristics and functions of osteoblasts, and this method is an ideal way to culture primary osteoblasts.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Osteoblast, Cell Culture Techniques, Pancreatin, Collagenases, Tissue

CLC Number:

R318

Ding Xiang-ying1, Cai Jing-wei2, Pan Ji-ming1, Liang Min2. Culture of rat primary osteoblasts using enzymatic digestion combined with tissue explant method[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(12): 1833-1837.

Figures/Tables 1

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