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人鼻中隔软骨细胞在无血清培养基中的生长及生长因子产生

Growth and growth factor production by human nasal septal chondrocytes in serum-free media.

作者信息

Kita Magdalena, Hanasono Matthew M, Mikulec Anthony A, Pollard Jeffrey D, Kadleck James M, Koch R James

机构信息

Koch Laboratory, Division of Otolaryngology-Head and Neck Surgery, Stanford University Medical Center, Stanford, California, USA.

出版信息

Am J Rhinol. 2006 Sep-Oct;20(5):489-95. doi: 10.2500/ajr.2006.20.2927.

Abstract

BACKGROUND

Tissue-engineered human cartilage offers vast possibilities as a source of graft implant material for reconstructive surgery. Serum-supplemented growth media is successful in supporting chondrocyte proliferation in vitro. Serum, however, contains exogenous growth factors that hamper the identification and quantification of growth factors autogenously produced by chondrocytes. We explore the possibility of using a commercially available serum-free medium UltraCULTURE as an alternative to modified Webber's medium (MWM), the standard media used in chondrocyte cell culture.

METHODS

Human nasal septal chondrocytes were grown in UltraCULTURE containing various concentrations of basic fibroblast growth factor (bFGF; 0, 1, 10, and 100 ng/mL) with or without insulin-like growth factor and compared with chondrocytes grown in MWM. Growth curves and transforming growth factor (TGF) beta 1 production were analyzed.

RESULTS

We found no differences in the ability to sustain cell viability in culture between the two base media types. We also found no statistically significant differences in TGF-beta 1 production by chondrocytes grown in either system. Finally, there were no statistically significant differences in chondrocyte proliferation between cultures supplemented with bFGF at 10 and 100 ng/mL.

CONCLUSION

UltraCULTURE media is a cost-effective, serum-free alternative to standard media with compatible growth characteristics. It offers specific advantages over standard serum-containing media for the precise measurement of autogenous growth factor production by cultured chondrocytes. Furthermore, UltraCULTURE's serum-free environment would be ideal for safely producing tissue-engineered cartilage grafts.

摘要

背景

组织工程化人软骨作为重建手术中移植植入材料的来源具有广阔的前景。补充血清的生长培养基在体外成功支持软骨细胞增殖。然而,血清含有外源性生长因子,这会妨碍对软骨细胞自身产生的生长因子的鉴定和定量。我们探讨了使用市售无血清培养基UltraCULTURE替代改良韦氏培养基(MWM,软骨细胞培养中使用的标准培养基)的可能性。

方法

将人鼻中隔软骨细胞接种于含有不同浓度碱性成纤维细胞生长因子(bFGF;0、1、10和100 ng/mL)且添加或不添加胰岛素样生长因子的UltraCULTURE培养基中培养,并与接种于MWM培养基中的软骨细胞进行比较。分析生长曲线和转化生长因子(TGF)β1的产生情况。

结果

我们发现两种基础培养基在维持培养细胞活力的能力上没有差异。我们还发现,在这两种培养体系中生长的软骨细胞产生TGF-β1的量没有统计学上的显著差异。最后,添加10和100 ng/mL bFGF的培养物中软骨细胞的增殖没有统计学上的显著差异。

结论

UltraCULTURE培养基是一种具有成本效益的无血清替代标准培养基,具有兼容的生长特性。与标准含血清培养基相比,它在精确测量培养软骨细胞自身生长因子产生方面具有特定优势。此外,UltraCULTURE的无血清环境对于安全生产组织工程化软骨移植物将是理想的。

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