S for their therapeutic effects have not been completely revealed. One possible explanation is that the transplanted stem cells generate cells function well as Gracillin normal hepatocytes do. However, the percentage of engraftment and graft survival after cell transplantation remains disappointing [2,16]. Another explanation is the indirect paracrine effects initiated in the damaged liver after stem cell transplant [14,17]. Some soluble factors such as cytokines or chemokines may have been secreted in order to facilitate the process of damage repair and liver regeneration. One of the CXCR3-related chemokines, the interferon-c-inducible protein 10 (IP-10), has been regarded as a marker of inflammatory damage. Besides, IP-10 expression was shown to correlate with the degree of liver inflammation, necrosis and fibrosis [18,19,20]. Recently it was found that IP-10 could modulate either positively or negatively the repair and the regeneration process in different forms of liver injuries [21,22,23,24]. Thus, we proposed that IP-10 may have an important regulatory role in the cell-base therapy for acute liver injury. At present, it remains unclear whether or not IP10 is involved in and regulates the recovery process of the injured liver after stem cell transplantation. In this study, we induced iPS to differentiate into hepatocytes in vitro. These cells were referred as iPS-derived hepatocyte-like cells (iHL), which functionally resemble primary hepatocytes. Then we investigated the effects of iPS and iHL on acute toxininduced liver injury and explored the possible underlying paracrine-mediated mechanism. Here, we showed that transplanted iPS increased the expression of IP-10 in injured liver to facilitate damage repair and promote liver regeneration.percentages of positive cells were highest in iPS group when compared to the control and iHL groups (Fig. 2A). Besides, significant numbers of fluorescent cells were detected in liver, spleen, lung, and bone marrow by flow-cytometry analysis (Fig. 2B Table S2). The majority of the DiI-labeled iPS was localized in the liver and spleen and the mean percentages were 2.66 and 4.74 , respectively. This implied that the iPS may function in liver through a direct or an indirect route. Additionally, no iPSinduced teratoma was detected in the study mice during a 6month observation period (Fig. S3).IPS Increased Hepatic IP-10 Expression 15857111 in Injured LiverThe iPS-induced cytokines changes in the liver were evaluated by cytokine array (Fig. 3A). Among all the cytokines tested, IP-10 and MIG were upregulated by 7- and 6-folds in liver tissues respectively. Further study showed that the mRNA expression of IP-10 and MIG significantly increased at 24 h post-injury (Fig. 3B). In contrast, the expression of iTAC, which 24786787 belong to the same cytokine family as IP-10 and MIG, decreased (Fig. 3B). At 48 h post-injury, the levels of IP-10 and MIG decreased, but the expression of IP-10 in the iPS group remained significantly order Lecirelin higher than that in the CCl4 group without iPS treatment (p,0.05). At protein levels, results from ELISA and Western blot analysis demonstrated that there was a significant increase of hepatic IP-10 by iPS at 24 h post-injury (Fig. 3C).IPS and Hepatocytes as the Cellular Sources of IP-Next we tested if iPS can secret IP-10 directly and be the source of IP-10 in vivo. The iPS was found to secrete IP-10 into culture medium at concentration about 14 pg/ml per 30,000 cells; while compatible number of hepatocytes (.S for their therapeutic effects have not been completely revealed. One possible explanation is that the transplanted stem cells generate cells function well as normal hepatocytes do. However, the percentage of engraftment and graft survival after cell transplantation remains disappointing [2,16]. Another explanation is the indirect paracrine effects initiated in the damaged liver after stem cell transplant [14,17]. Some soluble factors such as cytokines or chemokines may have been secreted in order to facilitate the process of damage repair and liver regeneration. One of the CXCR3-related chemokines, the interferon-c-inducible protein 10 (IP-10), has been regarded as a marker of inflammatory damage. Besides, IP-10 expression was shown to correlate with the degree of liver inflammation, necrosis and fibrosis [18,19,20]. Recently it was found that IP-10 could modulate either positively or negatively the repair and the regeneration process in different forms of liver injuries [21,22,23,24]. Thus, we proposed that IP-10 may have an important regulatory role in the cell-base therapy for acute liver injury. At present, it remains unclear whether or not IP10 is involved in and regulates the recovery process of the injured liver after stem cell transplantation. In this study, we induced iPS to differentiate into hepatocytes in vitro. These cells were referred as iPS-derived hepatocyte-like cells (iHL), which functionally resemble primary hepatocytes. Then we investigated the effects of iPS and iHL on acute toxininduced liver injury and explored the possible underlying paracrine-mediated mechanism. Here, we showed that transplanted iPS increased the expression of IP-10 in injured liver to facilitate damage repair and promote liver regeneration.percentages of positive cells were highest in iPS group when compared to the control and iHL groups (Fig. 2A). Besides, significant numbers of fluorescent cells were detected in liver, spleen, lung, and bone marrow by flow-cytometry analysis (Fig. 2B Table S2). The majority of the DiI-labeled iPS was localized in the liver and spleen and the mean percentages were 2.66 and 4.74 , respectively. This implied that the iPS may function in liver through a direct or an indirect route. Additionally, no iPSinduced teratoma was detected in the study mice during a 6month observation period (Fig. S3).IPS Increased Hepatic IP-10 Expression 15857111 in Injured LiverThe iPS-induced cytokines changes in the liver were evaluated by cytokine array (Fig. 3A). Among all the cytokines tested, IP-10 and MIG were upregulated by 7- and 6-folds in liver tissues respectively. Further study showed that the mRNA expression of IP-10 and MIG significantly increased at 24 h post-injury (Fig. 3B). In contrast, the expression of iTAC, which 24786787 belong to the same cytokine family as IP-10 and MIG, decreased (Fig. 3B). At 48 h post-injury, the levels of IP-10 and MIG decreased, but the expression of IP-10 in the iPS group remained significantly higher than that in the CCl4 group without iPS treatment (p,0.05). At protein levels, results from ELISA and Western blot analysis demonstrated that there was a significant increase of hepatic IP-10 by iPS at 24 h post-injury (Fig. 3C).IPS and Hepatocytes as the Cellular Sources of IP-Next we tested if iPS can secret IP-10 directly and be the source of IP-10 in vivo. The iPS was found to secrete IP-10 into culture medium at concentration about 14 pg/ml per 30,000 cells; while compatible number of hepatocytes (.
