Engineering Human Mesenchymal Stem Cells to Release Adenosine Using miRNA Technology
Adenosine is an important modulator of metabolic activity with powerful tissue- and cell-protective functions. Adenosine kinase (ADK), the major adenosine-regulating enzyme, is critical to adapt its intra- and extra-cellular levels in response to environmental changes. Lentiviral RNAi-mediated down-regulation of ADK in human mesenchymal stem cells (hMSCs) has therefore been considered an effective tool for engineering therapeutically effective adenosine-releasing cell grafts that could constitute patient-identical autologous implants for clinical application. We constructed lentiviral vectors that coexpress miRNA directed against ADK and an emerald green fluorescent protein (EmGFP) reporter gene. Following lentiviral transduction of hMSCs, we demonstrated up to 80% down-regulation of ADK and 98% transduction efficiency. Transduced hMSCs continued to express EmGFP after 4–6 consecutive passages and EmGFP-positive hMSC grafts survived in the hippocampal fissure of mouse brains and provided efficient adenosine-dependent neuroprotection in a mouse model of seizure-induced cell loss.
- 常用细胞凋亡检测方法(二)
- Identification of Oocyte Progenitor Cells in the Zebrafish Ovary
- Promotion of Duplex and Triplex DNA Formation by Polycation Comb-Type Copolymers
- Computer-Assisted Interpretation of Triacylglycerols Mass Spectra
- Gel-Based and Gel-Free Proteomic Technologies
- 动物细胞培养实验室必备仪器
- 内质网(endoplasmic reticulum,ER)
- 体外荧光法检测核内体早期动力学
- 联会复合体
- 植物体内可溶性糖含量的测定(蒽酮法)