Identification of cytokines involved in hepatic differentiation of mBM-MSCs under liver-injury conditions

AIM: To identify the key cytokines involved in hepatic differentiation of mouse bone marrow mesenchymal stem cells (mBM-MSCs) under liver-injury conditions. METHODS: Abdominal injection of CCl4 was adopted to duplicate a mouse acute liver injury model. Global gene expression analysis was performed to evaluate the potential genes involved in hepatic commitment under liver-injury conditions. The cytokines involved in hepatic differentiation of mBM-MSCs was function-ally examined by depletion experiment using specifi c antibodies, followed by rescue experiment and direct inducing assay. The hepatic differentiation was characterized by the expression of hepatic lineage genes and proteins, as well as functional features. RESULTS: Cytokines potentially participating in hepatic fate commitment under liver-injury conditions were initially measured by microarray. Among the up-regulated genes determined, 18 cytokines known to closely relate to liver growth, repair and development, were selected for further identif ication. The f ibroblast growth factor-4 (FGF-4), hepatocyte growth factor (HGF) and oncostatin M (OSM) were fi nally found to be involved in hepatic differentiation of mBM-MSCs under liver-injury conditions. Hepatic differentiation could be dramatically decreased after removing FGF-4, HGF and OSM from the liver-injury conditioned medium, and could be rescued by supplementing these cytokines. The FGF-4, HGF and OSM play different roles in the hepatic differentiation of mBM-MSCs, in which FGF-4 and HGF are essential for the initiation of hepatic differentiation, while OSM is critical for the maturation of hepatocytes. CONCLUSION: FGF-4, HGF and OSM are the key cytokines involved in the liver-injury conditioned medium for the hepatic differentiation of mBM-MSCs.

Cytokines Expression in Lymphocytes From Rats With Allergic Asthma Induced by Pleurotus sapidus besidiospores

Allergic asthma caused by mushroom spores (Pleurotus sapidus besidiospores) is a common health problem among mushroom-cultivating workers in China. An animal model of allergic asthma through the challenge of Pleurotus sapidus besidiospores in primed rats was developed for investigating the role of cytokines in the pathogenesis of the disease. In the study a series of related cytokines and their receptors, including their activity and mRNA levels of spleen lymphocytes isolated from asthmatic rats, were measured. Determined by 3H-TdR incorporation assay and NAG microcolorimetric assay, Con A-induced spleen lymphocyte proliferation and IL-2 activity in culture supernatants of spleen lymphocytes from 7-day challengedrats with allergic asthma increased significantly by 261% and 208%, respectively, as compared with those in the control. Cytokines and their receptor expression at mRNA levels were determined by RNA/cDNA hybridization, using (α-32P-dCTP radiolabeled cDNA probes for different cytokines and their receptors in vitro. The results showed that mRNA expression of IL-4, GM-CSF, IL-6, IL-2 and IL-2R, except IL-6R, in lymphocytes of 7-day-challenged asthma-suffering rats, increased significantly by 54%, 45%, 170%, 83% and 76%, respectively. In 2-day challenged-rats, mRNA levels of IL-4, IL-6 and IL-2 increased by 37%,58% and 125%, respectively, whereas mRNA leve1s of GM-CSF, IL-2R and IL-6R remained unchanged. Thus, the experimental results suggested a significant increase in TH2type cytokines in the pathogenesis of Pleurotus sapidus besidiospore-induced allergic asthma and IL-4 may play an essential role.

Effects of Various Factors on Proliferation Capacity and Cytotoxicity of Induced CIK of Patients with Tumor In Vitro

Objective： To evaluate effects of different factors on proliferation capacity and cytotoxicity of induced cytokine induced killer （CIK） cells in vitro, so as to provide experimental basis for cell therapy of CIK in tumor. Methods： Cytometry and MTT assay were used in detecting proliferation capacity and cytotoxicity of CIK. Results： The proliferation capacity and cytotoxicity of CIK in vitro were stronger in the group of low age and serum free medium than in the group of advanced age or serum medium. And the proliferation of CIK in vitro increased with time during a certain period of incubation. Furthermore, CIK had equal cytotoxicity to various tumor cells. Conclusion： Various factors might influence the proliferation capacity and cytotoxicity of CIK of tumor patients in vitro.