Stem cells are a kind of self-renewal and pluripotency cell population. Stem cells can be divided into embryonic stem cells and adult stem cells according to its origin. Liver stem cells belong to adult stem cells, which can be further divided into liver derived stem cells such as oval cells (OVs) and non- liver derived stem cells such as bone marrow hematopoietic stem cells and mesenchymal stem cells.
Recently, stem cells transplantation has achieved initial results in acute or chronic liver disease, but its pros and cons have been still in constant debate. HSCs located in the space of Disse are liver stromal cells which of great significance be involved in the liver’s physiological and pathological process. Previous studies have shown that HSCs activated in the acute or selleckchem chronic liver disease, transdifferentiated into myofibroblasts, secreted a mass of collagens and extracellular matrix, which
seriously damaged liver function and metabolism yet its normal morphology ever changed. Not until now, there has been reported numerously about HSCs but rarely refered to its further biological function or embryonic origin, which has been remained unknown. Therefore, we design our research as follows: 1. Isolation and identification of HSCs. Aquired target cells from rat liver and identified whether they were HSCs by a serial of experiments. 2. Detection of HSCs’ stem cell markers. Select stem cell markers of HSCs’ probably embryonic origin through RT-PCR and ICC. 3. Differentiation of HSCs into hepatocyte-like cells. Observed differentiation of HSCs transformed into hepatocyte-like
cells through Ixazomib mw cytokines induction in vitro. To sum up, we illustrated that: 1. Primary HSCs expressed selleck some stem cell markers. 2. HSCs could differentiate into hepatocyte-like cells after cytokines induction in vitro. To prove that HSCs might possess stem cell characteristic, and HSCs might be a population of stem cells/progentiors in liver. Methods: Method1. Isolation of primary HSCs of rat. SD rat weighted approximate 450–500 g, isolated HSCs from two step include primary liver perfusion combined with isolated liver perfusion and one step only consist of primary liver perfusion separately, then aquired target cells from density gradient centrifuge via medium 60% percoll.2. Identification of rat HSCs. Identificated of HSCs’ morphology, 328 nm autofluorescence, lipid droplets and specific cell markers.3. Detection of stem cell marker from HSCs. Dectated HSCs stem cell markers by RT-PCR and ICC.4. Cytokines inducted HSCs differentiated into hepatocyte-like cells. Two groups, control group cultured 24 h without cytokines, while the experimental group cultured within bFGF 20 ug/L, FGF4 20 ug/L, HGF 20 ug/L, IL-6 1 ug/L for first 3 days, then followed only FGF4 20 ug/L for another 4 days.5. Detected induced HSCs before and after whether expressed hepatocyte specific markers’ gene and protein by Real time PCR and ICC.6.