Goldberg A J, Lee D A, Bader D L, Bentley G
Interdisciplinary Research Centre, Institute of Orthopaedics, Royal Free and University College Medical School, Stanmore, Middlesex, England, UK.
J Bone Joint Surg Br. 2005 Jan;87(1):128-34.
An increasing number of patients are treated by autologous chondrocyte implantation (ACI). This study tests the hypothesis that culture within a defined chondrogenic medium containing TGF-beta enhances the re-expression of a chondrocytic phenotype and the subsequent production of cartilaginous extracellular matrix by human chondrocytes used in ACI. Chondrocytes surplus to clinical requirements for ACI from 24 patients were pelleted and cultured in either DMEM (Dulbecco's modified eagles medium)/ITS+Premix/TGF-beta1 or DMEM/10%FCS (fetal calf serum) and were subsequently analysed biochemically and morphologically. Pellets cultured in DMEM/ITS+/TGF-beta1 stained positively for type-II collagen, while those maintained in DMEM/10%FCS expressed type-I collagen. The pellets cultured in DMEM/ITS+/TGF-beta1 were larger and contained significantly greater amounts of DNA and glycosaminoglycans. This study suggests that the use of a defined medium containing TGF-beta is necessary to induce the re-expression of a differentiated chondrocytic phenotype and the subsequent stimulation of glycosaminoglycan and type-II collagen production by human monolayer expanded chondrocytes.
越来越多的患者接受自体软骨细胞植入(ACI)治疗。本研究检验了以下假设:在含有转化生长因子-β(TGF-β)的特定软骨形成培养基中培养,可增强用于ACI的人软骨细胞软骨细胞表型的重新表达以及随后软骨细胞外基质的产生。将24例患者ACI临床需求剩余的软骨细胞制成细胞团,分别在DMEM(杜氏改良伊格尔培养基)/ITS +预混物/TGF-β1或DMEM/10%胎牛血清(FCS)中培养,随后进行生化和形态学分析。在DMEM/ITS +/TGF-β1中培养的细胞团Ⅱ型胶原染色呈阳性,而在DMEM/10%FCS中培养的细胞团表达Ⅰ型胶原。在DMEM/ITS +/TGF-β1中培养的细胞团更大,且含有显著更多的DNA和糖胺聚糖。本研究表明,使用含有TGF-β的特定培养基对于诱导人单层扩增软骨细胞分化软骨细胞表型的重新表达以及随后刺激糖胺聚糖和Ⅱ型胶原的产生是必要的。