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人半月板细胞:原代培养的特性及在组织工程中的应用

Human meniscus cell: characterization of the primary culture and use for tissue engineering.

作者信息

Nakata K, Shino K, Hamada M, Mae T, Miyama T, Shinjo H, Horibe S, Tada K, Ochi T, Yoshikawa H

机构信息

Department of Orthopaedic Sports Medicine, Kansai Rosai Hospital, Hyogo, Japan.

出版信息

Clin Orthop Relat Res. 2001 Oct(391 Suppl):S208-18.

Abstract

Human meniscus cells from 47 surgically excised menisci were grown in primary culture. Cell proliferation and morphologic features were evaluated in three different culture media. Human meniscus cells showed three distinguishable cell types in monolayer culture: elongated fibroblastlike cells, polygonal cells, and small round chondrocytelike cells. These cells proliferated in Dulbecco's modified Eagle's medium, but by Day 7, elongated fibroblastlike cells became predominant. Cells did not proliferate in Ham's nutrient mixture-F-12. In a mixture of Ham's nutrient mixture-F-12 and Dulbecco's modified Eagle's medium, cells proliferated, maintaining their morphologic features and their ability to express messenger ribonucleic acids for aggrecan and Types I, II, and III collagen. Hyaluronan enhanced cellular proliferation without altering morphologic features or chondroitin sulfate production. Cultured human meniscus cells attached to a porous collagen sponge after cell seeding. Gene transfer was successful and an introduced gene was expressed by the cells, indicating that human meniscus cells can undergo gene manipulation. The finding that cells collected from small surgical specimens of human meniscus could be cultured, propagated, and seeded onto a collagen scaffold holds promise for the development of a cell-based, tissue engineered collagen meniscus.

摘要

对47个手术切除的人半月板中的细胞进行原代培养。在三种不同的培养基中评估细胞增殖和形态学特征。人半月板细胞在单层培养中表现出三种可区分的细胞类型:细长的成纤维细胞样细胞、多边形细胞和小圆形软骨细胞样细胞。这些细胞在杜氏改良 Eagle 培养基中增殖,但到第7天,细长的成纤维细胞样细胞占主导。细胞在Ham's营养混合液-F-12中不增殖。在Ham's营养混合液-F-12和杜氏改良 Eagle 培养基的混合物中,细胞增殖,保持其形态学特征以及表达聚集蛋白聚糖和I、II、III型胶原蛋白信使核糖核酸的能力。透明质酸增强细胞增殖,而不改变形态学特征或硫酸软骨素的产生。细胞接种后,培养的人半月板细胞附着在多孔胶原海绵上。基因转移成功,导入的基因由细胞表达,表明人半月板细胞可进行基因操作。从人半月板小手术标本中收集的细胞能够培养、增殖并接种到胶原支架上,这一发现为基于细胞的组织工程化胶原半月板的开发带来了希望。

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