In Vitro Hepatic Trans-Differentiation of Human Mesenchymal Stem Cells Using Sera from Congestive/Ischemic Liver during Cardiac Failure

Abstract

Cellular therapy for end-stage liver failures using human mesenchymal stem cells (hMSCs)-derived hepatocytes is a potential alternative to liver transplantation. Hepatic trans-differentiation of hMSCs is routinely accomplished by induction with commercially available recombinant growth factors, which is of limited clinical applications. In the present study, we have evaluated the potential of sera from cardiac-failure-associated congestive/ischemic liver patients for hepatic trans-differentiation of hMSCs. Results from such experiments were confirmed through morphological changes and expression of hepatocyte-specific markers at molecular and cellular level. Furthermore, the process of mesenchymal-to-epithelial transition during hepatic trans-differentiation of hMSCs was confirmed by elevated expression of E-Cadherin and down-regulation of Snail. The functionality of hMSCs-derived hepatocytes was validated by various liver function tests such as albumin synthesis, urea release, glycogen accumulation and presence of a drug inducible cytochrome P450 system. Based on these findings, we conclude that sera from congestive/ischemic liver during cardiac failure support a liver specific microenvironment for effective hepatic trans-differentiation of hMSCs in vitro.

A Patient‐Inspired Ex Vivo Liver Tissue Engineering Approach with Autologous Mesenchymal Stem Cells and Hepatogenic Serum

“…This infers that natural‐origin hepatogenic factors present in patient sera at physiological concentration is sufficient enough to drive the mesenchymal‐to‐epithelial transition during hepatic specification. In our previous studies, we have proved the phenomenon of mesenchymal‐to‐epithelial transition of BMSCs during hepatic trans‐differentiation using sera from cardiac failure patients with ischemic/congestive liver dysfunction, as confirmed by overexpression of E‐cadherin and downregulation of snail protein . There are reports that factors released from cholestatic rat liver possibly contribute to hepatic priming of bone marrow stem cells .…”

“…Cell Proliferation Assay on Fibrous Meshes in Presence of Hepatogenic Sera : We previously reported that 10% jaundiced sera was cytotoxic and 5% sera supported suitable BMSCs proliferation . In the present study, patient‐derived BMSCs (of passage 2) were seeded (at 5000 cells cm −2 ) on previously UV‐sterilized coverslips containing electrospun meshes of either PLLA or PLLA/gelatin in DMEM–LG containing either 5% FBS (Hyclone) or patient‐derived hepatogenic sera (HS) supplemented with 1% antibiotic and anti‐mycotic solution.…”

“…Collection of Hepatogenic Sera : In a previous work, we have reported that HGF‐enriched sera collected from patients with cardiac‐failure‐associated congestive/ischemic liver dysfunction (secondary hyperbilirubinemia) have significant hepatogenic role on human BMSCs. Accordingly, such sera was collected from patients ( n = 18, mean age group: 58 ± 14, male: 11, Type AB) who attended the biochemistry unit of International Centre for Cardiothoracic and Vascular Diseases, Frontier Lifeline Hospital, Chennai, India and pooled together (mean serum HGF concentration, 9.87 ± 2.3 ng mL −1 ).…”

“…It is known that during heart failure, both circulatory and myocardial concentrations of HGF is elevated significantly, which is shown to have cardioprotective role . Recently, we have reported that during cardiac‐failure‐associated secondary ischemic/congestive liver dysfunction, there is increased level of serum HGF, which could induce hepatic commitment of MSCs successfully …”

Targeting c-kit receptor in neuroblastomas and colorectal cancers using stem cell factor (SCF)-based recombinant bacterial toxins

“…Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood using Ficoll density gradient separation method. Human bone marrow cells previously isolated and preserved as described by Dilip et al (Bishi et al 2014) were used for this experiment. Cells were maintained in Iscove modified Dulbecco medium (IMDM), with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin (Pen Strep), and were cultured in a humidified atmosphere at 5 % CO 2 and 37°C.…”