Volume 26, Issue 4 (2023)
mjms 2023, 26(4): 33-45 |
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Malaki M, Tiraihi T, Ghorbani-Anarkooli M, Babaei A. Differential expression of growth and differentiation genes in murine parotid gland-derived stem cells cultured on three-dimensional alginate hydrogel. mjms 2023; 26 (4) :33-45
URL: http://mjms.modares.ac.ir/article-30-77452-en.html
URL: http://mjms.modares.ac.ir/article-30-77452-en.html
1- Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University
2- Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University ,takialtr@modares.ac.ir
2- Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University ,
Abstract: (504 Views)
Introduction: We aimed to compare the gene expression of parotid gland derived stem cell in a three (3D) alginate hydrogel culture with that of a two-dimensional one (2D).
Materials and methods: Five rats were sacrificed and the parotid glands were removed and cultured in DMEM/F12 medium supplemented with 15% FBS. The cells were characterized by flow cytometry and immunocytochemistry for evaluating the expression of genes. The cells were encapsulated in alginate hydrogel, then the differentiation was compared with that of a 2D culture. qRT-PCR was performed in order to evaluate the expression of Amy1, Cldn3, Cidn4, Ki67, Cyclin D1, Dpt, Meox2, Aquaporin 5, Pparg, Bpifa2e and Tp63 genes.
Results: The harvested cells immunoreacted with CD90, CD44, and CD29, however, the immunophenotyping of CD45 and CD34 were negative. The immunocytochemistry results showed that they were strongly immunostained with K-7 and E-cadherin, but less with K-14. In the 3D culture, the cells differentiated into organoid bodies with round shape, there was duct-like structure extended from one pole. The qRT-PCR in the 3D culture showed increase in the expression of Amy1, Ki67, aquaporin5, Pparg, Bpifa2e and Tp63 genes compared to 2D culture. In contrast Cldn3, Cldn4, cyclin D1, Dpt and Meox2 genes were strongly expressed in the 2D culture.
Conclusion: The results of flow cytometry and immunocytochemistry confirmed the properties of the isolated cells were parotid gland-derived mesenchymal stem cells. They differentiated into organoid body in the 3D culture using alginate as scaffold which expressed parotid gland differentiation genes.
Materials and methods: Five rats were sacrificed and the parotid glands were removed and cultured in DMEM/F12 medium supplemented with 15% FBS. The cells were characterized by flow cytometry and immunocytochemistry for evaluating the expression of genes. The cells were encapsulated in alginate hydrogel, then the differentiation was compared with that of a 2D culture. qRT-PCR was performed in order to evaluate the expression of Amy1, Cldn3, Cidn4, Ki67, Cyclin D1, Dpt, Meox2, Aquaporin 5, Pparg, Bpifa2e and Tp63 genes.
Results: The harvested cells immunoreacted with CD90, CD44, and CD29, however, the immunophenotyping of CD45 and CD34 were negative. The immunocytochemistry results showed that they were strongly immunostained with K-7 and E-cadherin, but less with K-14. In the 3D culture, the cells differentiated into organoid bodies with round shape, there was duct-like structure extended from one pole. The qRT-PCR in the 3D culture showed increase in the expression of Amy1, Ki67, aquaporin5, Pparg, Bpifa2e and Tp63 genes compared to 2D culture. In contrast Cldn3, Cldn4, cyclin D1, Dpt and Meox2 genes were strongly expressed in the 2D culture.
Conclusion: The results of flow cytometry and immunocytochemistry confirmed the properties of the isolated cells were parotid gland-derived mesenchymal stem cells. They differentiated into organoid body in the 3D culture using alginate as scaffold which expressed parotid gland differentiation genes.
Article Type: Original Research |
Subject:
Neuroscience (General)
Received: 2024/10/12 | Accepted: 2024/10/16
Received: 2024/10/12 | Accepted: 2024/10/16
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