软骨细胞体外增殖去分化的拉曼光谱分析

引用本文: 金璐頔, 徐晶晶, 张勇, 余跃洲, 刘畅, 赵东平, 叶安培. 软骨细胞体外增殖去分化的拉曼光谱分析[J]. 物理化学学报, 2017, 33(12): 2446-2453. doi: 10.3866/PKU.WHXB201706133 shu

Citation: JIN Lu-Di, XU Jing-Jing, ZHANG Yong, YU Yue-Zhou, LIU Chang, ZHAO Dong-Ping, YE An-Pei. Raman Spectroscopic Analysis of Chondrocyte Dedifferentiation during in vitro Proliferation[J]. Acta Physico-Chimica Sinica, 2017, 33(12): 2446-2453. doi: 10.3866/PKU.WHXB201706133 shu

软骨细胞体外增殖去分化的拉曼光谱分析

金璐頔 ^1, ,
徐晶晶 ^2, ,
张勇 ^3, ,
余跃洲 ^2, ,
刘畅 ^3, ,
赵东平 ^3, ,
叶安培 ^1,3,

1.
北京大学前沿交叉学科研究院, 北京 100871;
2.
北京大学-清华大学生命科学联合中心, 北京 100871;
3.
北京大学信息科学技术学院, 纳米器件物理与化学教育部重点实验室, 北京 100871
基金项目:
国家自然科学基金（U1636110），科技部“科技支撑计划”（2012BAF14B14）及北京大学“医学-信息”交叉研究种子基金（2014-MI-19）资助项目

摘要: 基于显微拉曼光谱技术，对组织工程的软骨种子细胞在传代增殖过程中的去分化进行单细胞分析。首先，对体外单层培养的第1-4代（P1-P4）大鼠软骨细胞样本进行了单细胞拉曼光谱检测，由此识别出软骨细胞中各种碱基、糖基、氨基酸等主要物质分子结构的特征峰集合。随后，分析拉曼光谱中若干重点特征峰强度随细胞传代次数的变化，发现软骨细胞体外增殖过程中核酸（789、1094、1576 cm^-1）含量降低、II型胶原（特异组分为羟脯氨酸，1207 cm^-1）和蛋白聚糖（特异组分为糖胺聚糖，1042、1063、1126、1160 cm^-1）合成下降、脂质（1304 cm^-1）及磷酸盐（957 cm^-1）含量增加等分子水平变化，从而在活体单细胞层次初步揭示了去分化引起软骨细胞增殖变缓、分泌减弱、形态纤维化等现象的分子机制。

关键词:

English

Raman Spectroscopic Analysis of Chondrocyte Dedifferentiation during in vitro Proliferation

1.
Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, P. R. China;
2.
Peking-Tsinghua Center for Life Sciences, Beijing 100871, P. R. China;
3.
Key Laboratory for the Physics & Chemistry of Nano-devices, School of Electronics Engineering & Computer Science, Peking University, Beijing 100871, P. R. China
Received Date: 02 May 2017
Revised Date: 05 June 2017
Fund Project:
The project was supported by the National Natural Science Foundation of China (U1636110), the National Key Technologies R&D Program of China (2012BAF14B14), and the Medicine-Informatics Interdisciplinary Project of Peking University, China (2014-MI-19).

Abstract: Seeded chondrocytes play a crucial role in current cartilage tissue engineering, yet both the quality and quantity of these cells could be impaired owing to cell dedifferentiation during in vitro proliferation. Here, we used micro-Raman spectroscopy to investigate changes in cellular components upon monolayer culturing of primary rat chondrocytes through multiple passages. Based on the average spectral profiles, we detected a series of Raman peaks and recognized related radicals such as nucleobases, pyranose rings, sulfate, tyrosine, proline, and amides at the single-chondrocyte level. Quantitative analysis of the Raman peak intensities showed that nucleic acids (at 789, 1094, 1576 cm^-1) decreased significantly from passage 1 (P1) to passage 4 (P4), whereas lipids (at 1304 cm^-1) and phosphate (at 957 cm^-1) increased significantly. Moreover, the syntheses of two major hyaline cartilage-associated proteins, aggrecan and type-2 collagen, were impeded, as indicated by the marked decline in the levels of their specific components (glycosaminoglycan at 1042, 1063, 1126, 1160 cm^-1, and hydroxyproline at 1207 cm^-1). Taken together, these features reveal the diminished propagation and secretion abilities of passaged chondrocytes needed for matrix-induced implantation, and shed light on the molecular mechanism of chondrocyte dedifferentiation.

Key words:

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软骨细胞体外增殖去分化的拉曼光谱分析

English

Raman Spectroscopic Analysis of Chondrocyte Dedifferentiation during in vitro Proliferation

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软骨细胞体外增殖去分化的拉曼光谱分析

English

Raman Spectroscopic Analysis of Chondrocyte Dedifferentiation during in vitro Proliferation

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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