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在受限体外组织培养系统中对人类椎间盘细胞功能的研究。

Studies of human intervertebral disc cell function in a constrained in vitro tissue culture system.

作者信息

Le Maitre Christine Lyn, Hoyland Judith Alison, Freemont Anthony J

机构信息

Laboratory Medicine Academic Group, University of Manchester, Manchester, United Kingdom.

出版信息

Spine (Phila Pa 1976). 2004 Jun 1;29(11):1187-95. doi: 10.1097/00007632-200406010-00006.

Abstract

STUDY DESIGN

This is a laboratory-based study examining a novel in vitro culture system for intervertebral disc tissue.

OBJECTIVES

Address the hypothesis that "the novel culture system will preserve intervertebral disc tissue matrix and cell function and prevent cellular apoptosis for periods up to 21 days."

SUMMARY OF BACKGROUND DATA

Studies of cell function in human intervertebral disc tissue are scarce. In vivo study of human intervertebral disc cells remains impracticable; in situ molecular biology in histologic sections lacks a dynamic dimension; and as for in vitro studies, cell culture often lacks physiologic relevance and explant cultures are subject to loss of tissue integrity and altered cell behavior. There is a biologic and therapeutic need for a satisfactory explant culture system for studying human intervertebral disc tissue in a controlled environment.

METHODS

Samples of human intervertebral disc tissue, obtained at surgery, were examined for a number of tissue and cell parameters immediately after excision (controls) and following culture of tissue samples either in a plastic ring or unconstrained in tissue culture medium for up to 3 weeks. Data were compared between cultured tissue and controls.

RESULTS

By comparison with control tissue, unconstrained explants swelled, tissue structure was disturbed, and there were profound changes in cell function. By contrast, tissue cultured in plastic rings maintained tissue structure, and after 3 weeks, the cellular parameters were the same as in controls.

CONCLUSIONS

This is the first reported system to preserve cell function of human discal explants for long periods in tissue culture. It will be a useful tool for a wide range of investigations of intervertebral disc biology that have not hitherto been possible.

摘要

研究设计

这是一项基于实验室的研究,旨在检验一种用于椎间盘组织的新型体外培养系统。

目的

验证“该新型培养系统能够在长达21天的时间内保持椎间盘组织基质和细胞功能,并防止细胞凋亡”这一假设。

背景资料总结

关于人类椎间盘组织细胞功能的研究较少。对人类椎间盘细胞进行体内研究仍然不可行;组织学切片中的原位分子生物学缺乏动态维度;至于体外研究,细胞培养往往缺乏生理相关性,而外植体培养容易出现组织完整性丧失和细胞行为改变的情况。在可控环境下研究人类椎间盘组织,需要一种令人满意的外植体培养系统,这在生物学和治疗学上都有需求。

方法

手术获取的人类椎间盘组织样本,在切除后立即(作为对照)以及将组织样本在塑料环中培养或在组织培养基中无约束培养长达3周后,检测一系列组织和细胞参数。对培养后的组织和对照进行数据比较。

结果

与对照组织相比,无约束的外植体肿胀,组织结构受到干扰,细胞功能发生深刻变化。相比之下,在塑料环中培养的组织保持了组织结构,3周后细胞参数与对照相同。

结论

这是首次报道的能够在组织培养中长期保持人类椎间盘外植体细胞功能的系统。它将成为迄今为止无法进行的广泛椎间盘生物学研究的有用工具。

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