Winter Anja, Breit Stephen, Parsch Dominik, Benz Karin, Steck Eric, Hauner Hans, Weber Renate M, Ewerbeck Volker, Richter Wiltrud
Orthopaedic Hospital/University of Heidelberg, Heidelberg, Germany.
Arthritis Rheum. 2003 Feb;48(2):418-29. doi: 10.1002/art.10767.
To compare the chondrogenic potential of human bone marrow-derived mesenchymal stem cells (BMSC) and adipose tissue-derived stromal cells (ATSC), because the availability of an unlimited cell source replacing human chondrocytes could be strongly beneficial for cell therapy, tissue engineering, in vitro drug screening, and development of new therapeutic options to enhance the regenerative capacity of human cartilage.
Quantitative gene expression of common cartilage and cell interaction molecules was analyzed using complementary DNA array technology and reverse transcription-polymerase chain reaction during optimization of cell differentiation, in order to achieve a molecular phenotype similar to that of chondrocytes in cartilage.
The multilineage potential of BMSC and ATSC was similar according to cell morphology and histology, but minor differences in marker gene expression occurred in diverse differentiation pathways. Although chondrogenic differentiation of BMSC and ATSC was indistinguishable in monolayer and remained partial, only BMSC responded (with improved chondrogenesis) to a shift to high-density 3-dimensional cell culture, and reached a gene expression profile highly homologous to that of osteoarthritic (OA) cartilage.
Hypertrophy of chondrocytes and high matrix-remodeling activity in differentiated BMSC spheroids and in OA cartilage may be the basis for the strong similarities in gene expression profiles between these samples. Differentiated stem cell spheroids represent an attractive tool for use in drug development and identification of drug targets in OA cartilage-like tissue outside the human body. However, optimization of differentiation protocols to achieve the phenotype of healthy chondrocytes is desired for cell therapy and tissue engineering approaches.
比较人骨髓间充质干细胞(BMSC)和脂肪组织来源的基质细胞(ATSC)的软骨形成潜力,因为获得一种无限的细胞来源来替代人软骨细胞可能对细胞治疗、组织工程、体外药物筛选以及开发增强人软骨再生能力的新治疗方案非常有益。
在细胞分化优化过程中,使用互补DNA阵列技术和逆转录-聚合酶链反应分析常见软骨和细胞相互作用分子的定量基因表达,以实现与软骨中软骨细胞相似的分子表型。
根据细胞形态学和组织学,BMSC和ATSC的多向分化潜能相似,但在不同的分化途径中标记基因表达存在微小差异。尽管BMSC和ATSC的软骨形成分化在单层培养中难以区分且仍不完全,但只有BMSC对向高密度三维细胞培养的转变有反应(软骨形成增强),并达到与骨关节炎(OA)软骨高度同源的基因表达谱。
分化的BMSC球体和OA软骨中软骨细胞的肥大和高基质重塑活性可能是这些样本基因表达谱强烈相似的基础。分化的干细胞球体是用于药物开发和识别体外OA软骨样组织中药物靶点的有吸引力的工具。然而,对于细胞治疗和组织工程方法,需要优化分化方案以实现健康软骨细胞的表型。