Voga Metka, Drnovsek Natasa, Novak Sasa, Majdic Gregor
Institute of Preclinical Sciences, Veterinary Faculty, University of Ljubljana, Ljubljana, Slovenia.
Department for Nanostructured Materials, Jozef Stefan Institute, Ljubljana, Slovenia.
J Tissue Eng. 2019 Mar 14;10:2041731419835056. doi: 10.1177/2041731419835056. eCollection 2019 Jan-Dec.
Under appropriate culture conditions, mesenchymal stem cells (MSC), also called more properly multipotent mesenchymal stromal cells (MMSC), can be induced toward differentiation into different cell lineages. In order to guide stem cell fate within an environment resembling the stem cell niche, different biomaterials are being developed. In the present study, we used silk fibroin (SF) as a biomaterial supporting the growth of MMSC and studied its effect on chondrogenesis of canine adipose-derived MMSC (cADMMSC). Adipose tissue was collected from nine privately owned dogs. MMSC were cultured on SF films and SF scaffolds in a standard cell culture medium. Cell morphology was evaluated by scanning electron microscopy (SEM). Chondrogenic differentiation was evaluated by alcian blue staining and mRNA expression of collagen type 1, collagen type 2, Sox9, and Aggrecan genes. cADMMSC cultured on SF films and SF scaffolds stained positive using alcian blue. SEM images revealed nodule-like structures with matrix vesicles and fibers resembling chondrogenic nodules. Gene expression of chondrogenic markers Sox9 and Aggrecan were statistically significantly upregulated in cADMMSC cultured on SF films in comparison to negative control cADMMSC. This result suggests that chondrogenesis of cADMMSC could occur when cells were grown on SF films in a standard cell culture medium without specific culture conditions, which were previously considered necessary for induction of chondrogenic differentiation.
在适当的培养条件下，间充质干细胞（MSC），更确切地称为多能间充质基质细胞（MMSC），可以被诱导分化为不同的细胞谱系。为了在类似于干细胞生态位的环境中引导干细胞命运，人们正在开发不同的生物材料。在本研究中，我们使用丝素蛋白（SF）作为支持MMSC生长的生物材料，并研究其对犬脂肪来源的MMSC（cADMMSC）软骨形成的影响。从9只私人饲养的犬只中采集脂肪组织。MMSC在标准细胞培养基中的SF膜和SF支架上培养。通过扫描电子显微镜（SEM）评估细胞形态。通过阿尔新蓝染色以及1型胶原、2型胶原、Sox9和聚集蛋白聚糖基因的mRNA表达评估软骨分化。在SF膜和SF支架上培养的cADMMSC经阿尔新蓝染色呈阳性。SEM图像显示出具有基质小泡和类似软骨结节纤维的结节样结构。与阴性对照cADMMSC相比，在SF膜上培养的cADMMSC中软骨形成标志物Sox9和聚集蛋白聚糖的基因表达在统计学上显著上调。该结果表明，当细胞在标准细胞培养基中的SF膜上生长时，cADMMSC的软骨形成可能会发生，而无需先前认为诱导软骨分化所必需的特定培养条件。
