Abstract
The treatment of neurodegenerative diseases is still a challenging grindstone in reconstructive surgeries and regenerative medicine. The retention of mesenchymal stem cells (MSCs) to retain remarkable properties of differentiating into motor neuron-like cells and Schwann cells can prove to be effective in repairing disorders. Moreover, the ultrafine electrospun nanofibers provide a favorable and conducive platform for proliferation and differentiation of MSCs. The development of new 3D culture methods with electrospun scaffolds that closely mimic the physiological niche of cells will help us to understand the functional benefits of MSCs in regeneration process. This article highlights the protocols for isolation of MSCs from rat bone marrow and their subsequent culture on nanofiber scaffolds. Furthermore, this chapter summarizes the various procedures including isolation of the MSCs, their seeding on electrospun nanofibrous scaffolds, and their proliferation and differentiation into neural lineage upon appropriate induction. The materials and preparation of various reagents used at different steps of the protocol are also summarized in detail.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ansari S, Diniz IM, Chen C, Sarrion P, Tamayol A, Wu BM, Moshaverinia A (2017) Human periodontal ligament-and Gingiva-derived mesenchymal stem cells promote nerve regeneration when encapsulated in alginate/hyaluronic acid 3D scaffold. Adv Healthc Mater 6(24):1700670
Rinker B, Zoldos J, Weber RV, Ko J, Thayer W, Greenberg J, Leversedge FJ, Safa B, Buncke G (2017) Use of processed nerve allografts to repair nerve injuries greater than 25 mm in the hand. Ann Plast Surg 78(6S):S292–S295
Cooney DS, Wimmers EG, Ibrahim Z, Grahammer J, Christensen JM, Brat GA, Wu LW, Sarhane KA, Lopez J, Wallner C (2016) Mesenchymal stem cells enhance nerve regeneration in a rat sciatic nerve repair and hindlimb transplant model. Sci Rep 6:31306
Sofi HS, Ashraf R, Beigh MA, Sheikh FA (2018) Scaffolds fabricated from natural polymers/composites by electrospinning for bone tissue regeneration. In: Cutting-edge enabling technologies for regenerative medicine. Springer, Singapore, pp 49–78
Sankar S, Sharma CS, Rath SN, Ramakrishna S (2017) Electrospun fibers for recruitment and differentiation of stem cells in regenerative medicine. Biotechnol J 12(12):1700263
Shall G, Menosky M, Decker S, Nethala P, Welchko R, Leveque X, Lu M, Sandstrom M, Hochgeschwender U, Rossignol J (2018) Effects of passage number and differentiation protocol on the generation of dopaminergic neurons from rat bone marrow-derived mesenchymal stem cells. Int J Mol Sci 19(3):720
Wang HS, Hung SC, Peng ST, Huang CC, Wei HM, Guo YJ, Fu YS, Lai MC, Chen CC (2004) Mesenchymal stem cells in the Wharton’s jelly of the human umbilical cord. Stem Cells 22(7):1330–1337
Xu Y, Zhang Z, Chen X, Li R, Li D, Feng S (2016) A silk fibroin/collagen nerve scaffold seeded with a co-culture of schwann cells and adipose-derived stem cells for sciatic nerve regeneration. PLoS One 11(1):e0147184
Chamberlain G, Fox J, Ashton B, Middleton J (2007) Concise review: mesenchymal stem cells: their phenotype, differentiation capacity, immunological features, and potential for homing. Stem Cells 25(11):2739–2749
Kwon A, Kim Y, Kim M, Kim J, Choi H, Jekarl DW, Lee S, Kim JM, Shin J-C, Park IY (2016) Tissue-specific differentiation potency of mesenchymal stromal cells from perinatal tissues. Sci Rep 6:23544
Jung JP, Bache-Wiig MK, Provenzano PP, Ogle BM (2016) Heterogeneous differentiation of human mesenchymal stem cells in 3D extracellular matrix composites. Biores Open Access 5(1):37–48
Agmon G, Christman KL (2016) Controlling stem cell behavior with decellularized extracellular matrix scaffolds. Curr Opin Solid State Mater Sci 20(4):193–201
Sofi HS, Ashraf R, Khan AH, Beigh MA, Majeed S, Sheikh FA (2018) Reconstructing nanofibers from natural polymers using surface functionalization approaches for applications in tissue engineering, drug delivery and biosensing devices. Mater Sci Eng C 94(1):1102–1124
Ashraf R, Sofi HS, Beigh MA, Majeed S, Arjamand S, Sheikh FA (2018) Prospects of natural polymeric scaffolds in peripheral nerve tissue-regeneration. In: Novel biomaterials for regenerative medicine. Springer, Singapore, pp 501–525
Ashraf R, Sofi HS, Malik A, Beigh MA, Hamid R, Sheikh FA (2019) Recent trends in the fabrication of starch nanofibers: electrospinning and non-electrospinning routes and their applications in biotechnology. Appl Biochem Biotechnol 187(1):47–74
Theron A, Zussman E, Yarin A (2001) Electrostatic field-assisted alignment of electrospun nanofibres. Nanotechnology 12(3):384
Zhang K, Zheng H, Liang S, Gao C (2016) Aligned PLLA nanofibrous scaffolds coated with graphene oxide for promoting neural cell growth. Acta Biomater 37:131–142
Wang X, Ding B, Li B (2013) Biomimetic electrospun nanofibrous structures for tissue engineering. Mater Today 16(6):229–241
Bonino CA, Efimenko K, Jeong SI, Krebs MD, Alsberg E, Khan SA (2012) Three-dimensional electrospun alginate nanofiber mats via tailored charge repulsions. Small 8(12):1928–1936
Dumas V, Guignandon A, Vico L, Mauclair C, Zapata X, Linossier MT, Bouleftour W, Granier J, Peyroche S, Dumas J-C (2015) Femtosecond laser nano/micro patterning of titanium influences mesenchymal stem cell adhesion and commitment. Biomed Mater 10(5):055002
Bagher Z, Azami M, Ebrahimi-Barough S, Mirzadeh H, Solouk A, Soleimani M, Ai J, Nourani MR, Joghataei MT (2016) Differentiation of Wharton’s jelly-derived mesenchymal stem cells into motor neuron-like cells on three-dimensional collagen-grafted nanofibers. Mol Neurobiol 53(4):2397–2408
Acknowledgments
Dr. Faheem A. Sheikh acknowledges the financial support given by Nano Mission Council, Department of Science and Technology, under grant number SR/NM/NM-1038/2016, and Science and Engineering Research Board (SERB) under grant number ECR/2016/001429.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media New York
About this protocol
Cite this protocol
Ashraf, R., Sofi, H.S., Sheikh, F.A. (2019). Experimental Protocol of MSC Differentiation into Neural Lineage for Nerve Tissue Regeneration Using Polymeric Scaffolds. In: Turksen, K. (eds) Stem Cell Nanotechnology. Methods in Molecular Biology, vol 2125. Humana, New York, NY. https://doi.org/10.1007/7651_2019_229
Download citation
DOI: https://doi.org/10.1007/7651_2019_229
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0359-8
Online ISBN: 978-1-0716-0360-4
eBook Packages: Springer Protocols