Study On Transplantation Of Adult Stem Cells To Rat Lower Esophagus

Background:Gastroesophageal reflux disease (GERD) has a high prevalence of 51% in Western countries. A dysfunction of the lower esophageal sphincter of unknown origin is the main etiology. The investigation in adult in Beijing and Shanghai showed the prevalence rate is 5.77%, but the practical prevalence rate is much more than it. Besides the classic symptoms (heartburn and regurgitation), GERD can also cause the extraesophageal disorders, which can impact on the qualfty-of -life of the patients. And Barrett's esophagus, one of GERD complications, is the risk factor of esophageal adenocarcinoma.Principal regimens for patients with GERD have included (1)general measures; (2)pharmacologic therapy, which is the mainstay of healing and prevention of recurrence of GERD;(3)surgical treatment;(4)endoscopic technic. Howerer, all of the above methods have deficiency and none of them can cure GERD completely.Objective:To explore the transplantation of adult stem cells (muscle satellite cells and hone marrow mesenchymal stem cells) as a new method curing gastroesophageal reflux disease, we determined the feasiblity and safety injecting adult stem cells into lower esophagus of rats.Methods:1. The skeletal muscle satellite cells ( SC) and the bone marrow mesenchymal stem cells (MSC ) were cultured in vitro, which were isolated from the tibialis anterior muscles and the whole hind limbs of Wistar rats respectively.2. After labelled with 5-bromc-2-deoxyuridine (BrdU), the satellite cells were injected into the lamina muscularis of lower esophagus of autologous rats, and the mesenchymal stem cells were injected into the same position of allogenic rats.3. Tissue of lower esophagus was harvested after 1 and 4 weeks, sectioned, stained for HE and immunohistochemistry to value the development and differentiation of the injected cells.Results:1. At least 1X104 satellite cells were isolated from each of rat tibialis anterior muscle weighted more than 0.5 grams. And there were 80% SCs in culture flask after purification by differential adhesion.2. More than 5 X106 /ml mesenchymal stem cells were isolated from the whole hind limb bone marrow of a rat.3. One and four week after cell transplantation, a lot of injected cells were observed in the lamina muscularis of lower esophagus in both of groups, which had differentiated into esophageal skeletal muscle cells. Local inflammation was not found obviously.4. In the lamina muscularis of lower esophagus in SC group, the cells labelled by BrdU at the fourth week were more than those at the first week (P<0.01) .5. There were a slight increase of cells labelled by BrdU at the fourth weekcompared with the first week in MSC group (p>0.05) .6. The significant increase of cells labelled by BrdU was detected in SC group compared with MSC group at the fourth week (P<0.05) . However, there were no significant differences in SC and MSC groups at the first week (p>0.05) .Conclusions:We proved the feasibility injecting muscle satellite cells and the mesenchymal stem cells into the lower esophagus and the persistence of injected cells. Transplantation of adult stem cells may be an attractive potential regimens to cure gastroesophageal reflux disease.