| 骨细胞分离培养及其与成骨细胞鉴别比较的实验研究 |
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Received:June 20, 2012
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| 作者 | Author | 单位 | Unit | E-Mail |
| 汤小康 |
TANG Xiao-kang |
浙江中医药大学第一临床医学院, 浙江 杭州 310053 |
The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China |
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| 程婉 |
CHENG Wan |
浙江中医药大学第一临床医学院, 浙江 杭州 310053 |
The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China |
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| 许兵 |
XU Bing |
浙江中医药大学第一临床医学院, 浙江 杭州 310053 |
The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China |
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| 应航 |
YING Hang |
浙江中医药大学第一临床医学院, 浙江 杭州 310053 |
The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China |
yh@zjtm.net |
| 童培建 |
TONG Pei-jian |
浙江中医药大学第一临床医学院, 浙江 杭州 310053
浙江省中医院, 浙江 杭州 310006 |
The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China |
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| 肖鲁伟 |
XIAO Lu-wei |
浙江中医药大学第一临床医学院, 浙江 杭州 310053 |
The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China |
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| 期刊信息：《中国骨伤》2013年26卷，第3期，第227-231页 |
| DOI：10.3969/j.issn.1003-0034.2013.03.014 |
| 基金项目:浙江省自然基金(编号;Y2110928);浙江省中医药科学研究基金计划(编号;2010ZA020 |
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目的：建立比较稳定的骨细胞实验室分离培养方法,并将其部分生物学特性与实验室培养的成骨细胞进行比较研究,明确两者区别。
方法：采用序列酶消化法,分别从3只3 d SD乳鼠骨骼中分离培养骨细胞和成骨细胞,培养24 h后进行细胞形态学观察。第1代细胞用碱性磷酸酶试剂盒采用重氮盐法(改良Kaplow氏法)染色,采用免疫细胞化学法对细胞的骨钙素(BGP)染色,测定碱性磷酸酶并计算其活性。
结果：骨细胞多呈星状或树枝状,且有很多的突触;成骨细胞呈长梭形,有少量的突触。骨细胞碱性磷酸酶染色,胞浆内cAKP颗粒不明显;成骨细胞碱性磷酸酶染色,胞浆内可见众多cAKP颗粒。骨细胞BGP染色阳性明显,成骨细胞BGP染色阳性不如骨细胞明显。ALP在骨细胞中分泌较成骨细胞低,且有统计学差异。
结论：实验室条件下能培养出骨细胞,该类细胞和成骨细胞有明显区别。 |
| [关键词]:骨细胞 成骨细胞 细胞分离 |
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| Experimental study on the isolated culture of osteocytes and identification of osteoblasts in rats |
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Abstract:
Objective: To establish a more stable method to isolate osteocytes in vitro,and then to find the differences with osteoblast biological characteristics.
Methods: Osteocytes and osteoblasts were isolated from the bone tissue of 3-day-old rats using sequential collagenase digestion. The cells were identified through cell morphology after 24 hours later. Alkaline phosphatase(ALP) kit was used to stain the first generation cells by Kaplow-way,the bone gla protein(BGP) of the cells were stained by immunocytochemitry. Measured ALP and computed its activity.
Results: Osteocytes and osteoblasts showed obviously differences in cell morphology. Osteocytes were star-shaped or dendrite-shaped within more dendrites,while osteoblasts were spindle-shaped with short dendrites. Osteocytes were negative for ALP,but osteoblasts were positive; Osteocytes were more positive for BGP,and osteoblasts were less positive. The secretion of ALP in osteocytes was lower than that of osteoblasts.
Conclusion: Osteocytes can be isolated and cultured in vitro. These characteristics of osteocytes are apparently difference with those of osteoblasts. |
| KEYWORDS:Osteocytes Osteoblasts Cell separation |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 汤小康,程婉,许兵,应航,童培建,肖鲁伟.骨细胞分离培养及其与成骨细胞鉴别比较的实验研究[J].中国骨伤,2013,26(3):227~231 |
| 英文格式: | TANG Xiao-kang,CHENG Wan,XU Bing,YING Hang,TONG Pei-jian,XIAO Lu-wei.Experimental study on the isolated culture of osteocytes and identification of osteoblasts in rats[J].zhongguo gu shang / China J Orthop Trauma ,2013,26(3):227~231 |
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