Study on the Viability and Proliferation of Single Mesenchymal Stem Cell Encapsulated in Alginate and Alginate-PLL Using Microfluidic Approach

Document Type : Original Research

Authors

Institute of Life Science, Tarbiat Modares University, Tehran, Iran

Abstract
Stem cells are characterized by their capacity for self-renewal and their ability to differentiate into specific cell types under the influence of their microenvironment. It is known that matrix chemistry controls stem cell differentiation. Single cell encapsulations of the Mesenchymal stem cells into a semi-permeable microgel, allows a greater control of the stem cell fate. In this study, a chip for single-cell encapsulation was designed and fabricated using microfluidic technology. By using microfluidic chip, human bone marrow mesenchymal stem cells (hBMSCs) are encapsulated inside alginate and alginate-poly-l lysine (PLL) microgels. The results of long-term viability of MSCs inside alginate-PLL microgels, shows a significant increase compared to alginate microgels. Mesenchymal stem cell proliferation in alginate-PLL microgels also increased significantly on days 14 and 21. It seems that PLL improves cell adhesion and function by creating a positively charged matrix. Microscopic studies indicate that the morphology of the cells inside the microgels is spherical. However, the average diameter and volume of cells in microgels containing PLL are smaller than others, which indicates more proliferation and space limitation inside the microgels. Therefore, single cell alginate-PLL microgels provide a suitable substrate in clinical studies for tissue engineering, organ transplantation and cell therapy.

Keywords

Subjects


1. Zhang, L. and T.J. Webster, Nanotechnology and nanomaterials: promises for improved tissue regeneration. Nano today, 2009. 4(1): p. 66-80.
2. Bokhari, M.A., et al., The enhancement of osteoblast growth and differentiation in vitro on a peptide hydrogel—polyHIPE polymer hybrid material. Biomaterials, 2005. 26(25): p. 5198-5208.
3. Ermis, M., E. Antmen, and V. Hasirci, Micro and Nanofabrication methods to control cell-substrate interactions and cell behavior: A review from the tissue engineering perspective. Bioactive materials, 2018. 3(3): p. 355-369.
4. Habanjar, O., et al., 3D Cell Culture Systems: Tumor Application, Advantages, and Disadvantages. International Journal of Molecular Sciences, 2021. 22(22): p. 12200.
5. Gasperini, L., J.F. Mano, and R.L. Reis, Natural polymers for the microencapsulation of cells. Journal of the royal society Interface, 2014. 11(100): p. 20140817.
6. Park, H., et al., Effect of swelling ratio of injectable hydrogel composites on chondrogenic differentiation of encapsulated rabbit marrow mesenchymal stem cells in vitro. Biomacromolecules, 2009. 10(3): p. 541-546.
7. Temenoff, J.S., et al., In vitro osteogenic differentiation of marrow stromal cells encapsulated in biodegradable hydrogels. Journal of Biomedical Materials Research Part A: An Official Journal of The Society for Biomaterials, The Japanese Society for Biomaterials, and The Australian Society for Biomaterials and the Korean Society for Biomaterials, 2004. 70(2): p. 235-244.
8. Zhang, M.W., et al., Adapting biodegradable oligo (poly (ethylene glycol) fumarate) hydrogels for pigment epithelial cell encapsulation and lens regeneration. Tissue Engineering Part C: Methods, 2010. 16(2): p. 261-267.
9. Tendulkar, S., et al., A three-dimensional microfluidic approach to scaling up microencapsulation of cells. Biomedical microdevices, 2012. 14(3): p. 461-469.
10. Aguilar, L.M.C., et al., Formation of alginate microspheres prepared by optimized microfluidics parameters for high encapsulation of bioactive molecules. Journal of Colloid and Interface Science, 2021. 587: p. 240-251.
11. Rogan, H., F. Ilagan, and F. Yang, Comparing single cell versus pellet encapsulation of mesenchymal stem cells in three-dimensional hydrogels for cartilage regeneration. Tissue Engineering Part A, 2019. 25(19-20): p. 1404-1412.
12. An, C., et al., Continuous microfluidic encapsulation of single mesenchymal stem cells using alginate microgels as injectable fillers for bone regeneration. Acta Biomaterialia, 2020. 111: p. 181-196.
13. Utech, S., et al., Microfluidic generation of monodisperse, structurally homogeneous alginate microgels for cell encapsulation and 3D cell culture. Advanced healthcare materials, 2015. 4(11): p. 1628-1633.
14. Mohajeri, M., et al., Cell encapsulation in alginate-based microgels using droplet microfluidics; a review on gelation methods and applications. Biomedical Physics & Engineering Express, 2022. 8(2): p. 022001.
15. Huebsch, N., et al., Harnessing traction-mediated manipulation of the cell/matrix interface to control stem-cell fate. Nature materials, 2010. 9(6): p. 518-526.
16. Li, F., et al., Microfluidic encapsulation of human mesenchymal stem cells for articular cartilage tissue regeneration. ACS applied materials & interfaces, 2017. 9(10): p. 8589-8601.
17. Sarker, B., et al., Evaluation of fibroblasts adhesion and proliferation on alginate-gelatin crosslinked hydrogel. PloS one, 2014. 9(9): p. e107952.
18. Sarker, B., et al., Alginate-based hydrogels with improved adhesive properties for cell encapsulation. International journal of biological macromolecules, 2015. 78: p. 72-78.
19. De Pascalis, C. and S. Etienne-Manneville, Single and collective cell migration: the mechanics of adhesions. Molecular biology of the cell, 2017. 28(14): p. 1833-1846.
20. Wang, Y., Programmable hydrogels. Biomaterials, 2018. 178: p. 663-680.