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Heparin/Collagen 3D Scaffold Accelerates Hepatocyte Differentiation of Wharton’s Jelly-Derived Mesenchymal Stem Cells

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Both mature and stem cell-derived hepatocytes lost their phenotype and functionality under conventional culture conditions. However, the 3D scaffolds containing the main extracellular matrix constitutions, such as heparin, may provide appropriate microenvironment for hepatocytes to be functional. The current study aimed to investigate the efficacy of the differentiation capability of hepatocytes derived from human Wharton’s jelly mesenchymal stem cells (WJ-MSCs) in 3D heparinized scaffold. In this case, the human WJ-MSCs were cultured on the heparinized and non-heparinized 2D collagen gels or within 3D scaffolds in the presence of hepatogenic medium. Immunostaining was performed for anti-alpha fetoprotein, cytokeratin-18 and -19 antibodies. RT-PCR was performed for detection of hepatic nuclear factor-4 (HNF-4), albumin, cytokeratin-18 and -19, glucose-6-phosphatase (G6P), c-met and Cyp2B. The results indicated that hepatogenic media induced the cells to express early liver-specific markers including HNF4, albumin, cytokeratin-18 and 19 in all conditions. The cells cultured on both heparinized culture conditions expressed late liver-specific markers such as G6P and Cyp2B as well. Besides, the hepatocytes differentiated in 3D heparinized scaffolds stored more glycogen that indicated they were more functional. Non-heparinized 2D gel was the superior condition for cholangiocyte differentiation as indicated by higher levels of cytokeratin 19 expression. In conclusion, the heparinized 3D scaffolds provided a microenvironment to mimic Disse space. Therefore, 3D heparinized collagen scaffold can be suggested as a good vehicle for hepatocyte differentiation.

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Acknowledgements

The authors wish to thank the Iran National Science Foundation: INSF for offering the Grant No. 91003812 and also Ms. M. Sani and M. Rozitalab for technical supports and Dr. N. Shokrpour for English improvement. All experiments were performed in the laboratory of stem cell research and tissue engineering lab of Anatomy department, Shiraz University of Medical Sciences.

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Authors and Affiliations

  1. Department of Biology, Kharazmi University, No. 43, South Mofatteh Ave, Tehran, 15712-14911, Iran

    Fatemeh Aleahmad & Mahnaz Azarnia

  2. Biochemistry Department, Shiraz University of Medical Sciences, Mehr Intersection, Qasr-Dasht st., District 1, Shiraz, Fars, 71345, Iran

    Sepideh Ebrahimi

  3. Tissue Engineering Lab, Anatomy Department, Shiraz Medical School, Shiraz University of Medical Sciences, Mehr Intersection, Qasr-Dasht st., District 1, Shiraz, Fars, 71345, Iran

    Mahin Salmannezhad, Mojtaba Hoseini & Tahereh Talaei-Khozani

  4. Institute for Cancer Research, Shiraz University of Medical Sciences, Mehr Intersection, Qasr-Dasht st., District 1, Shiraz, Fars, 71345, Iran

    Mansooreh Jaberipour

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  1. Fatemeh Aleahmad
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  2. Sepideh Ebrahimi
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Correspondence to Tahereh Talaei-Khozani.

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Ethical statement

The experimental design was approved by ethic committee of Shiraz University of Medical Sciences (IR.SUMS.REC.1395.S803).

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Aleahmad, F., Ebrahimi, S., Salmannezhad, M. et al. Heparin/Collagen 3D Scaffold Accelerates Hepatocyte Differentiation of Wharton’s Jelly-Derived Mesenchymal Stem Cells. Tissue Eng Regen Med 14, 443–452 (2017). https://doi.org/10.1007/s13770-017-0048-z

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  • DOI: https://doi.org/10.1007/s13770-017-0048-z

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