Multipotent stromal cells stimulate liver regeneration by influencing the macrophage polarization in rat

作     者：Andrey Elchaninov Timur Fatkhudinov Natalia Usman Irina Arutyunyan Andrey Makarov Anastasia Lokhonina Irina Eremina Viktor Surovtsev Dmitry Goldshtein Galina Bolshakova Valeria Glinkina Gennady Sukhikh 

作者机构：National Medical Research Center for ObstetricsGynecology and Perinatology named after Academician V.I.Kulakov of Ministry of Healthcare of Russian FederationMoscow 117997Russia Peoples Friendship University of Russia(RUDN University)Moscow 117198Russia Scientific Research Institute of Human MorphologyMoscow 117418Russia Pirogov Russian National Research Medical UniversityMinistry of Healthcare of the Russian FederationMoscow 117997Russia Research Center of Medical GeneticsMoscow 115478Russia 

出 版 物：《World Journal of Hepatology》 (世界肝病学杂志（英文版）（电子版）)

年 卷 期：2018年第10卷第2期

页      面：287-296页

学科分类：1002[医学-临床医学] 10[医学] 

基　　金：Supported by Russian Science Foundation No.17-15-01419 

主　　题：Liver Regeneration Multipotent stromal cells Macrophages 

摘      要：AIM To investigate the influence of the umbilical cordderived multipotent stromal cells(MSCs) on recovery of the liver after the subtotal resection, that is, removal of 80% of the organ mass, a renowned model of the small-for-size liver remnant syndrome. METHODS The MSCs were obtained from the intervascular tissue of umbilical cords, dissected from rat fetuses, by the explant culture technique. The vital labeling of MSCs with РКН26 was carried out on the 3 rd passage. The subtotal resection was performed on male Sprague-Dawley rats. The experimental group animals received a transplant 106 MSCs infused into the spleen. Hepatocyte proliferation was assessed by counting of either mitotic figures or Ki67-positive cells in microscopic images. MSC differentiation was assessed with antibodies to hepatocyte-specific marker cytokeratin 18(CK18), cholangiocyte-specific protein CK19, smooth muscle cell-specific protein α-SMA, the endothelial cell marker CD31, or the active fibroblast marker FAPα. Total macrophages of the liver were selectively stained in cryosections incubated with antiCD68 antibodies(1:100, Abcam), while the M2 a and M2 c macrophage populations were selectively stained with anti-CD206 antibodies. Expression of interleukin and growth factor genes was evaluated with PCR-RT.RESULTS Intrasplenic allogeneic transplantation of the umbilical cord-derived multipotent stromal cells stimulates reparative processes within the residual liver tissue after subtotal resection(removal of 80% of the organ mass), as indicated by increased rates of hepatocyte proliferation and accelerated organ mass recovery. These effects may result from paracrine influence of the transplanted cells on the resident macrophage population of the liver. The transplantation favors polarization of macrophages to M2 phenotype(the M2-polarized macrophages specifically express CD206; they are known to suppress inflammation and support tissue repair). No differentiation of the transplanted cells into any of the liver cell types have been observed in the study.CONCLUSION We found no direct evidence for the paracrine effect of MSCs on liver regeneration after the subtotal liver resection in rats. However, the paracrine mechanism of the therapeutic activity of transplanted MSC is indirectly indicated by a decrease in the total number of CD68 + macrophages and an increase in the proportion of M2 pro-repair macrophages in the regenerating liver as compared to animals in which the transplantation was only mimicked.