Impact of tissue inhibitor of matrixmetalloprotease-1 (TIMP-1) protein on the proliferation and gene expression of rabbit Achilles tenocytes and rabbit adipose-derived stem cells in vitro

In order to know the effect of TIMP-1 on in vitro cell culture of rabbit Achilles tenocytes and rabbit adipose-derived stem cells, we assessed their proliferation with Alamar Blue® assay and furthermore report the effect on gene expression of Col1A1 (extracellular matrix marker collagen I), MKI67 (proliferation marker ki67), TNMD (tenomodulin) and ALP (alkaline phosphatase, a marker for early osteogenesis), respectively, the latter because TIMP-1 has been reported to affect stem cell fate towards osteogenic commitment. As reference gene, 18S was utilized.
These experiments have been performed at different concentrations of TIMP-1 supplementation; i.e. at 1, 10, and 100 ng/mL TIMP-1. The same experiment without TIMP-1 served as control.

为探究基质金属蛋白酶组织抑制剂-1（TIMP-1）对兔跟腱腱细胞及兔脂肪源性干细胞的体外细胞培养效应，本研究采用阿尔玛蓝（Alamar Blue®）检测法评估两类细胞的增殖能力，并进一步分别分析TIMP-1对Col1A1（细胞外基质标志物Ⅰ型胶原）、MKI67（增殖标志物ki67）、TNMD（腱调蛋白）以及ALP（碱性磷酸酶，早期成骨标志物）的基因表达影响——选择ALP作为检测指标之一，是因为已有研究证实TIMP-1可影响干细胞向成骨谱系分化。本次实验以18S作为内参基因。
本实验设置了不同浓度的TIMP-1干预组，即分别添加1、10、100 ng/mL的TIMP-1；同时设置不添加TIMP-1的对照组进行平行实验。