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3D Printing of Differentiated Bone Marrow Mesenchymal Cells as a New Method for Liver Tissue Engineering

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Three-dimensional-printed Bone marrow mesenchymal cells (BMSCs) after differentiation were used as in vitro seeds to synthesize minimal functional liver units. A model was developed for enabling three-dimensional (3D) regular seeding and culturing of differentiated bone marrow mesenchymal cells (dBMSCs) through the use of bioprinting technology to promote and maintain 3D liver tissue-like cellular morphology and cell-specific functionality in vitro with the addition of hydrogels. qPCR was used to detect the expression of ALB, HNF4a and AFP. The synthesis of glycogen was evaluated by periodic acid-Schiff staining. The expression of CK18 was observed under immunofluorescence after differentiation. The biocompatibility of the hydrogels was evaluated by MTT. The concentrations of GST, ALB and CYP2C9 in vitro were evaluated at 3 days, 7 days and 10 days. An in vivo experiment was performed by engrafting the printed tissue subcutaneously. The concentrations of ALB, CYP2C9 and GST in different engrafted tissues were quantified at different time points. The expressions of mRNA ALB, AFP and HNF4a in different groups were compared at 2 weeks and 4 weeks. From the differentiation process of BMSCs, we demonstrated that after several days of culturing, the mRNA concentrations of ALB, HNF4a and AFP changed significantly (p < 0.05). The marker of CK18 was expressed, and PSA staining was positive after differentiation. The biocompatibility of the 3D printing of hydrogels showed no significant difference from the 2D culture and mix culture. After the 3D printing, the concentrations of ALB, GST and CYP2C9 improved significantly after 3 days, 7 days and 10 days (p < 0.05). Liver tissue was formed after 4 weeks by 3D printing. The mRNAs of ALB, AFP and HNF4a were improved significantly in the printing culture group. The concentrations of ALB, GST and CYP2C9 improved significantly after 2 weeks and 4 weeks in in vivo experiments. Three-dimensional printing of dBMSCs can be used as a model of liver tissue engineering and in liver disease model research.
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Keywords: 3D PRINTING; BMSCS; LIVER REGENERATION

Document Type: Research Article

Publication date: August 1, 2016

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  • Journal of Biomaterials and Tissue Engineering (JBT) is an international peer-reviewed journal that covers all aspects of biomaterials, tissue engineering and regenerative medicine. The journal focuses on the broad spectrum of research topics including all types of biomaterials, their properties, bioimplants and medical devices, biofilms, bioimaging, BioMEMS/NEMS, biosensors, fibers, tissue scaffolds, tissue engineering and modeling, artificial organs, tissue interfaces, interactions between biomaterials, blood, cells, tissues, and organs, regenerative medicine and clinical performance.
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