Abstract
The injured or otherwise damaged cornea is healed by limbal stem cells (LSC). If the limbus where LSC reside is also damaged or nonfunctional, the cornea cannot heal properly and this defect leads to impaired vision that can result in blindness. The only way to treat total LSC deficiency is by transplantation of limbal tissue or a transfer of LSC. Recently, mesenchymal stem cells (MSC) have been shown as another promising source of stem cells for corneal healing and regeneration. Here, we describe a protocol for the use of polyamide 6/12 nanofiber scaffolds for the growth of MSC and LSC, and for their transfer onto a mechanically damaged ocular surface in the experimental mouse model.
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
Ma Y, Xu Y, Xiao Y, Yang W, Zhang C, Song E et al (2006) Reconstruction of chemically burned rat cornea surface by bone marrow-derived human mesenchymal stem cells. Stem Cells 24:315–321
Oh JY, Kim MK, Shin MS, Lee HJ, Ko JH, Wee WR et al (2008) The anti-inflammatory and anti-angeogenic role of mesenchymal stem cells in corneal wound healing Âfollowing chemical injury. Stem Cells 26:1047–1055
Zajicova A, Pokorna K, Lencova A, Krulova M, Svobodova E, KubÃnova S et al (2010) Treatment of ocular surface injuries by limbal and mesenchymal stem cells growing on nanofiber scaffolds. Cell Transplant 19:1281–1290
Rama P, Bonini S, Lambiase A, Golisano O, Paterna P, De Luca M et al (2001) Autologous fibrin-cultured limbal stem cells permanently restore the corneal surface of patients with total limbal stem deficiency. Transplantation 72:1478–1485
Schwab IR, Johnson NT, Harkim DG (2006) Inherent risks associated with manufacture of bioengineered ocular surface tissue. Arch Ophthalmol 124:1734–1740
Tsai RJ, Li LM, Chen JK (2000) Reconstruction of damaged cornea by transplantation of autologous limbal epithelial cells. N Eng J Med 343:86–93
Xie J, Willerth SM, Li X, Macewan MR, Rader A, Sakiyama-Elbert SE et al (2009) The differentiation of embryonic stem cells seeded on electrospun nanofibers into neural lineages. Biomaterials 30:354–362
Xin X, Hussain M, Mao JJ (2007) Continuing differentiation of human mesenchymal stem cells and induced chondrogenic and osteogenic lineages in electrospun PLGA nanofiber scaffold. Biomaterials 28:316–325
Shin YR, Chen CN, Tsai SW, Wang YJ, Lee OK (2006) Growth of mesenchymal stem cells on electrospun type I collagen nanofibers. Stem Cells 24:2391–2397
Nur-E-Kamal A, Ahmed I, Kamal J, Schindler M, Meiners S (2006) Three-dimensional nanofibrillar surfaces promote self-renewal in mouse embryonic stem cells. Stem Cells 24:426–433
Holan V, Chudickova M, Trosan P, Svobodova E, Krulova M, Kubinova S et al (2011) Cyclosporine A-loaded and stem cell-seeded electrospun nanofibers for cell-based therapy and local immunosuppression. J Control Release 156:406–412
Jirsak O, Sanetrnik OF, LukasD, Kotek K, Martinova L, Chaloupek J (2005) U. S. patent No. WO 205024101, 2005
Dubsky M, Kubinova S, Sirc J, Voska L, Svobodova J, Zajicek R et al (2012) Nanofibers prepared by needleless electrospinning technology as scaffolds for wound healing. J Mater Sci Mater Med 23:931–941
Krulova M, Pokorna K, Lencova A, Zajicova A, Fric J, Filipec M et al (2008) A rapid separation of two distinct populations of corneal epithelial cells with limbal stem cell characteristics in the mouse. Invest Ophthalmol Vis Sci 49:3903–3908
Xing X, Wang Y, Li B (2008) Nanofibers drawing and nanodevices assembly in poly (trimethylene terephtalate). Opt Express 16:10815–10822
Niece KL, Hartgerink JD, Donners JM, Stupp SI (2003) Self-assembly combining two bioactive peptide-amphiphile molecules into nanofibers by electrostatic attraction. J Am Chem Soc 125:7146–7147
Huang ZM, Zhang YZ, Kotaki M, Ramakrishna S (2003) A review on polymer nanofibers by electrospinning and their applications in nanocomposites. Compos Sci Technol 63:2223–2253
Svobodova E, Krulova M, Zajicova A, Pokorna K, Prochazkova J, Trosan P et al (2012) The role of mouse mesenchymal stem cells in differentiation of naive T-cells into anti-inflammatory regulatory T-cell or proinflammatory helper T-cell 17 population. Stem Cells Dev 21:901–910
Trosan P, Svobodova E, Chudickova M, Krulova M, Zajicova A, Holan V (2012) The key role of insulin-like growth factor I in Âlimbal stem cell differentiation and the corneal wound healing process. Stem Cells Dev 21:3341–3350
Acknowledgments
This work was supported by grants P304/11/0653 and P301/11/1568 from the Grant Agency of the Czech Republic, grant KAN200520804 from the Grant Agency of the Academy of Sciences, projects MSM0021620858 and SVV 265211 from the Ministry of Education of the Czech Republic, and project RVO 68378050 from the Academy of Sciences of the Czech Republic.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media New York
About this protocol
Cite this protocol
Holan, V., Javorkova, E., Trosan, P. (2013). The Growth and Delivery of Mesenchymal and Limbal Stem Cells Using Copolymer Polyamide 6/12 Nanofiber Scaffolds. In: Wright, B., Connon, C. (eds) Corneal Regenerative Medicine. Methods in Molecular Biology, vol 1014. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-432-6_13
Download citation
DOI: https://doi.org/10.1007/978-1-62703-432-6_13
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-431-9
Online ISBN: 978-1-62703-432-6
eBook Packages: Springer Protocols