: Significant difference between group A and group D at the same incubation time. gene-chip array and RT-qPCR. RESULTS More C3A cells attached to the plate made up of our serum-free medium than to those made up of HepatoZYME and DMEM/F12 at 24 h post-seeding. Bax inhibitor peptide P5 Both the viability and proliferation rate of C3A cells in sericin-based serum-free medium were superior to those of cells in HepatoZYME and DMEM/F12 ( 0.001). The content of albumin and urea in our serum-free medium was significantly higher than that in HepatoZYME and DMEM/F12 throughout the whole culture period ( 0.001) and was comparable to that in complete medium at day 3, 4, and 5. In part 2, cell viability and proliferation were greater in the presence of 2 mg/mL sericin ( 0.001), as was the proportion of cells in S phase (16.21% 0.98% 12.61% 0.90%, 0.01). Gene-chip array analysis indicated that this expression of were up-regulated by 2 mg/mL sericin, and RT-qPCR revealed that this expression of and was up-regulated by 2 mg/mL sericin ( 0.05). CONCLUSION We developed a novel hepatocyte serum-free medium. Sericin probably enhances cell attachment through the CCR6-Akt-JNK-NF-B pathway and promotes cell proliferation through CCR6-mediated activation of the ERK1/2-MAPK pathway. culture of C3A cells and applied an advanced method, gene-chip array, to Rabbit Polyclonal to PDGFRb (phospho-Tyr771) explore the effect of sericin around the hepatocyte transcriptome. We found that sericin probably enhanced cell attachment through the CCR6-Akt-JNK-NF-B pathway and promoted cell proliferation through CCR6-mediated activation of the ERK1/2-MAPK pathway. These findings inspired the following study around the mechanism by which sericin promotes cell attachment and proliferation. INTRODUCTION The bioartificial liver support system (BALSS) is usually a novel and ideal therapy for hepatic insufficiency, which can provide additional liver function for patients with acute liver injury and end-stage liver failure[1]. During the BALSS operation, hepatocytes in the bioreactor perform numerous functions such as albumin synthesis, ammonia removal, and bilirubin metabolism, which can decrease the symptoms of liver failure[2]. The BALSS is mainly composed of a hepatocyte culture module and an extracorporeal blood circulation device[3]. At the present time, the cells used in BALSS are mainly main porcine hepatocytes[4] and immortalized cells, such as HepG2 and C3A[5]. C3A is usually a human hepatocellular carcinoma cell collection, with high albumin production and excellent ability of ammonia removal. Therefore, C3A is usually selected as the hepatocyte in the extracorporeal liver assist device (ELAD), which has proven to be effective in liver support and biocompatible in patients in clinical trials[6]. Normally, culture of hepatocytes requires serum of animal-origin. However, the serum possesses several shortcomings, including immunogenicity, allergenicity and exposure to Bax inhibitor peptide P5 microorganisms[7]. During the operation of the BALSS, the hepatocyte culture medium is in contact with the patients plasma in the bioreactor, resulting in the potential for a variety of adverse reactions such as anaphylaxis and bacteremia. Therefore, serum-free medium suitable for hepatocyte culture in the BALSS has been needed Bax inhibitor peptide P5 within recent decades. However, few studies have focused on this topic. HepatoZYME-SFM, the most popular of all hepatocyte serum-free media, is usually a serum-free medium for the long-term maintenance of hepatocyte phenotypic expression including the active and inducible forms of cytochrome P450 and active phase II enzymes[8]. However, it is usually mainly used for serum-free main hepatocyte culture, and serum is required for the adherence of hepatocytes at the early stage of serum-free culture with HepatoZYME. Generally, serum-free medium comprises nutrients, growth factors, adherence-promoting factors, hormones, and trace elements. Advanced DMEM/F-12 (Dulbeccos Modified Eagle Medium/Hams F-12) is usually a widely used basal medium that allows the culture of mammalian cells with reduced Bax inhibitor peptide P5 (10-50 mL/L) fetal bovine serum (FBS) supplementation, so it is usually often selected as the basal medium of the serum-free medium. Growth factor is the key component of serum-free culture medium, as it promotes cell growth. Hepatocyte growth factor (HGF) is usually a key ligand that elicits G1/S progression of epithelial cells, including hepatocytes, by up-regulating cyclin-E1 the proline-mTOR pathway[9]. Epidermal growth factor (EGF) is not only a promoter of the growth of epithelial cells but also an important regulator that promotes CYP3A4 expression in hepatocytes[10]. Dexamethasone.