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Hindi Induction of hepatic regeneration in an experimental model using hepatocyte differentiated MSCs
Background and Objectives: Scaffolds are threedimensional
(3D) matrices that provide support for cells to attach, proliferate,
and differentiate, facilitating extracellular matrix formation. The
study aimed to examine the differentiation potential of Mesenchymal
stem cells (MSCs) into hepatocytes in 2D and 3D culture systems
to improve their in vitro differentiation, and test their functionality in
vivo.
Methods: MSCs were generated from umbilical cord blood.
Hepatogenic differentiation was induced on 2D and 3D cultures
and characterized by morphology, scanning electron microscopy,
immunocytochemistry and Gene expression. Albumin and
α-1 antitrypsin (AAT) in culture supernatants were measured.
Differentiated Cells were administered IV into a murine model of
carbon tetra (CCL4) induced liver cirrhosis which were divided into
3 groups, a) Pathological control group, b) and c) Groups treated
with hepatogenic differentiated MSCs cultured on 2D and 3D culture
system respectively. After 12 weeks of injection, liver pathology was
examined.
Results: The hepatogenic differentiated MSCs stained positively
for albumin, alpha fetoprotein (AFP), Heppar1, cytokeratin7, 18, and
OV6 with more mature cells, hexagonal in shape with central nuclei
forming large sheets in groups in 3D culture system. AAT secretion
and Indocyanine green uptake were significantly increased. in 3D
system. In experimental model, MSC-3D treated group exhibited
maximal restoration of liver architecture with absent septal fibrosis
and marked improvement of ALT, AST.
Conclusions: Both 3D and 2D culture system are effective
in functional hepatogenic differentiation from MSCs. In vivo
hepatogenic differentiation is more effective on 3D scaffold, with
better functional recovery.
