Expression Of Endothelial Nitric Oxide Synthase In Feline Mesenchymal Stromal Cells Mediated By Retrovirus

[Background and Objective]Achalasia is an idiopathic esophageal motor disorder characterized by an elevated pressure of lower esophageal sphincter (LES) and aperistalsis in the smooth muscle part of the esophageal body. The primary symptoms is dysphagia, which usually occurs with solids and liquids. It has been demonstrated that the smooth muscle of LES is normal in a state of contraction and relaxation, but the inhibitory NOS neurons which result in relaxation of LES in the myenteric nerve plexus decrease or are absent, and the stimulant cholinergic neurons which result in contraction of LES are in a relatively excited state. The treatment for achalasia includes drug treatment, endoscopic pneumatic balloon dilation, injection of botulinum toxin (BT) and surgical treatment (mostly laparoscopic). It is clear that all these therapies are palliative, and have their limitations respectively.Mesenchymal stromal cells (MSCs) are multipotential stem cells originated from mesoblast. They exist in the connective tissue and organ stroma and are abundant in bone marrows. In a certain condition they are capable of differentiating into all sorts of tissue cells. Because they are easily to be transfected by exogenous genes and can express exogenous genes steadily, MSCs are considered optimal target cells for tissue engineering, cell transplantation and gene therapy, and bone marrow stromal cells are becoming the hotspot of MSCs study fields.Retrovirus can not only transfer genes to proliferative cells effectively, but also integrate itself into the genome of the cells and do no harm to vectors. The aim of this experiment was to construct replication-deficient recombinant retrovirus carrying eNOS gene for gene transfer in MSCs and to measure the expression of eNOS, which could provide a new pathway for gene therapy in achalasia. [Methods]1. Construction of recombinant retrovirus pMSCVpuro-eNOS...