Hepatic differentiation of mouse bone marrow-derived mesenchymal stem cells using a novel 3D culture system

Abstract

The development of novel culture systems that mimic the in vivo microenvironment may be beneficial for inducing the differentiation of stem cells and promoting liver function. In the present study, spheroid cultures and decellularized liver scaffolds (DLSs) were utilized to obtain differentiated hepatocyte-like cells. Mouse bone marrow (BM)-derived mesenchymal stem cells (MSCs) self-aggregated into spheroids under low-attachment conditions and implanted into the DLSs via a negative pressure suction device. The Albp-ZsGreen adenoviral vector was utilized for real-time monitoring of hepatocyte-like cell differentiation. To detect the differentiation stages of the MSCs, immunostaining of hepatocyte markers and functional analysis was performed. Compared with traditional 2D monolayer induction, mouse BM-MSCs spheroids and DLSs in 3D culture generated greater yields of mature, differentiated hepatocytes. In conclusion, this 3D culture system may provide a strategy for generating hepatocyte-like cells for portable liver micro-organs, and aid clinical hepatocyte transplantation and liver tissue engineering.

Mesenchymal stem cell 3D encapsulation technologies for biomimetic microenvironment in tissue regeneration

“…In fact, cells show different physiological and morphological results in two-dimensional (2D) and 3D environments [10]. In particular, MSCs have better osteogenic [11], adipogenic [12], and hepatic [13, 14] differentiation behavior in the 3D environment. Moreover, MSCs show improved differentiation when they are co-cultured with other types of cells such as human umbilical vein endothelial cells (HUVECs) [15], osteoblasts [16], and hematopoietic stem/progenitor cells (HSPCs) [17] compared with those cultured alone in the 3D environment.…”

Therapeutic Potential of “Exosomes Derived Multiple Allogeneic Proteins Paracrine Signaling: Exosomes d-MAPPS” is Based on the Effects of Exosomes, Immunosuppressive and Trophic Factors

“…In addition to the cells of mesodermal origin (osteoblasts, chondroblasts, and adipocytes), MSCs are capable of generating neural cells, hepatocytes, alveolar epithelial cells, insulinproducing cells, cardiomyocytes, indicating their clinical application (7)(8)(9)(10)(11)(12)(13)(14). Several lines of evidence suggested that MSCs have capacity to differentiate into functional cardiomyocytes (9), hepatocytes (10,11), alveolar epithelial cells and lung precursor cells (13) contributing to the regeneration of injured myocardium, liver and lungs (12,(15)(16)(17). Additionally, in paracrine manner, through the production of immunomodulatory factors (indoleamine 2,3-dioxygenase (IDO), prostaglandin E2 (PGE2), nitric oxide (NO), transforming growth factor beta (TGF-β), interleukin (IL)-10, interleukin 1 receptor antagonist (IL-1Ra) and growth related oncogene (GRO), MSCs are able to suppress detrimental autoimmune response and to attenuate autoimmune and chronic, inflammatory diseases (3,18,19).…”

Augmenter of liver regeneration enhances cell proliferation through the microRNA‐26a/Akt/cyclin D1 pathway in hepatic cells

“…Mesenchymal stem cells are shown to play an important role in hepatic differentiation . During hepatic differentiation, there were significant changes in the expression of miRNAs in MSCs . We further investigated whether our observations described above hold true in human primary cells.…”

“…33 During hepatic differentiation, there were significant changes in the expression of miRNAs in MSCs. 34,35 We further investigated whether our observations described above hold true in human primary cells. Chronic liver diseases are characterized by high levels of circulating cytokines and growth factors, such as interferon-γ, IL-10, IL-4, IL-2, HGF, EGF, and TNF-α.…”