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The Use of Human Wharton’s Jelly Cells for Cochlear Tissue Engineering

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Auditory and Vestibular Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1427))

Abstract

Tissue engineering focuses on three primary components: stem cells, biomaterials, and growth factors. Together, the combination of these components is used to regrow and repair damaged tissues that normally do not regenerate easily on their own. Much attention has been focused on the use of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), due to their broad differentiation potential. However, ESCs and iPSCs require very detailed protocols to differentiate into target tissues, which are not always successful. Furthermore, procurement of ESCs is considered ethically controversial in some regions and procurement of iPSCs requires laborious transformation of adult tissues and characterization. However, mesenchymal stem cells are an adult stem cell population that are not ethically controversial and are readily available for procurement. Furthermore, mesenchymal stem cells exhibit the ability to differentiate into a variety of cell types arising from the mesoderm. In particular, human Wharton’s jelly cells (hWJCs) are mesenchymal-type stem cells found in umbilical cords that possess remarkable differentiation potential. hWJCs are a highly desirable stem cell population due to their abundance in supply, high proliferation rates, and ability to differentiate into multiple cell types arising from all three germ layers. hWJCs are used to generate several neurological phenotypes arising from the ectoderm and are considered for engineering mechanosensory hair cells found in the auditory complex. Here, we report the methods for isolating hWJCs from human umbilical cords and non-virally transfected for use in cochlear tissue engineering studies.

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References

  1. Chen J, Steit A (2013) Introduction of the inner ear: stepwise specification of otic fate from multipotent progenitors. Hear Res 297:3–12

    Article  PubMed  Google Scholar 

  2. Collado MS, Holt JR (2009) Can neurosphere production help restore inner ear transduction? Proc Natl Acad Sci U S A 106:8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Dhara SK, Gerwe BA, Majumder A, Dodla MC, Boyd NL, Machacek DW, Hasneen K, Stice SL (2009) Genetic manipulation of neural progenitors derived from human embryonic stem cells. Tissue Eng Part A 15:3621–3634

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Dhara SK, Majumder A, Dodla MC, Stice SL (2011) Nonviral gene delivery in neural progenitors derived from human pluripotent stem cells. Methods Mol Biol 767:343–354

    Article  CAS  PubMed  Google Scholar 

  5. Wang L, Ott L, Seshareddy K, Weiss ML, Detamore MS (2011) Musculoskeletal tissue engineering with human umbilical cord mesenchymal stromal cells. Regen Med 6:95–109

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Wataya T, Muguruma K, Sasai Y (2008) Human pluripotent stem cell and neural differentiation. Brain Nerve 60:1165–1172

    CAS  PubMed  Google Scholar 

  7. Koehler KR, Hashino E (2014) 3D mouse embryonic stem cell culture for generating inner ear organoids. Nat Protoc 9:1229–1244

    Article  CAS  PubMed  Google Scholar 

  8. Koehler KR, Mikosz AM, Molosh AI, Patel D, Hashino E (2013) Generation of inner ear sensory epithelia from pluripotent stem cells in 3D culture. Nature 500:217–221

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Longworth-Mills E, Koehler KR, Hashino E (2015) Generating inner ear organoids from mouse embryonic stem cells. Methods Mol Biol 1341:391–406

    Google Scholar 

  10. Ohnishi H, Skerleva D, Kitajiri S, Sakamoto T, Yamamoto N, Ito J, Nakagawa T (2015) Limited hair cell induction from human induced pluripotent stem cells using a simple stepwise method. Neurosci Lett 599:49–54

    Google Scholar 

  11. Yamahara K, Yamamoto N, Nakagawa T, Ito J (2015) Insulin-like growth factor 1: a novel treatment for the protection or regeneration of cochlear hair cells. Hear Res 330:2–9

    Google Scholar 

  12. Bancroft GN, Sikavitsas VI, van den Dolder J, Sheffield TL, Ambrose CG, Jansen JA, Mikos AG (2002) Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteoblasts in a dose-dependent manner. Proc Natl Acad Sci U S A 99:12600–12605

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Barry F, Boynton RE, Liu B, Murphy JM (2001) Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation-dependent gene expression of matrix components. Exp Cell Res 268:189–200

    Article  CAS  PubMed  Google Scholar 

  14. Romanov YA, Darevskaya AN, Merzlikina NV, Buravkova LB (2005) Mesenchymal stem cells from human bone marrow and adipose tissue: isolation, characterization, and differentiation potentialities. Bull Exp Biol Med 140:138–143

    Article  CAS  PubMed  Google Scholar 

  15. Rose RA, Jiang H, Wang X et al (2008) Bone marrow-derived mesenchymal stromal cells express cardiac-specific markers, retain the stromal phenotype, and do not become functional cardiomyocytes in vitro. Stem Cells 26:2884–2892

    Article  CAS  PubMed  Google Scholar 

  16. Bailey M, Wang L, Bode C, Mitchell K, Detamore MS (2007) Comparison of human umbilical cord matrix stem cells and temporomandibular joint condylar chondrocytes for tissue engineering temporomandibular joint condylar cartilage. Tissue Eng 13:2003–2010

    Article  CAS  PubMed  Google Scholar 

  17. Detamore MS (2013) Human umbilical cord mesenchymal stromal cells in regenerative medicine. Stem Cell Res Ther 4:142

    Article  PubMed  PubMed Central  Google Scholar 

  18. Devarajan K, Forrest ML, Detamore MS, Staecker H (2013) Adenovector-mediated gene delivery to human umbilical cord mesenchymal stromal cells induces inner ear cell phenotype. Cell Reprogram 15:43–54

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Mitchell KE, Weiss ML, Mitchell BM, Martin P, Davis D, Morales L, Helwig B, Beerenstrauch M, Abou-Easa K, Hildreth T, Troyer D, Medicetty S (2003) Matrix cells from Wharton’s jelly form neurons and glia. Stem Cells 21:50–60

    Article  CAS  PubMed  Google Scholar 

  20. Baksh D, Yao R, Tuan RS (2007) Comparison of proliferative and multilineage differentiation potential of human mesenchymal stem cells derived from umbilical cord and bone marrow. Stem Cells 25:1384–1392

    Article  CAS  PubMed  Google Scholar 

  21. Mellott AJ, Godsey ME, Shinogle HE, Moore DS, Forrest ML, Detamore MS (2014) Improving viability and transfection efficiency with human umbilical cord Wharton’s jelly cells through use of a ROCK inhibitor. Cell Reprogram 2:91–97

    Google Scholar 

  22. Mellott AJ, Devarajan K, Shinogle HE et al (2015) Nonviral reprogramming of human Wharton’s jelly cells reveals differences between ATOH1 homologues. Tissue Eng Part A 21:1795–1809

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Bustin SA, Mueller R (2005) Real-time reverse transcription PCR (qRT-PCR) and its potential use in clinical diagnosis. Clin Sci (Lond) 109:365–379

    Article  CAS  Google Scholar 

  24. Qiagen (2010) Critical factors for successful real-time PCR [E-Reader Version]. Accessed from https://www.qiagen.com/us/resources/resourcedetail?id=f7efb4f4-fbcf-4b25-9315-c4702414e8d6&lang=en

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Acknowledgments

This work was supported by NIH R01-AR056347 and the State of Kansas.

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Correspondence to Hinrich Staecker .

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Mellott, A.J., Detamore, M.S., Staecker, H. (2016). The Use of Human Wharton’s Jelly Cells for Cochlear Tissue Engineering. In: Sokolowski, B. (eds) Auditory and Vestibular Research. Methods in Molecular Biology, vol 1427. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3615-1_19

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  • DOI: https://doi.org/10.1007/978-1-4939-3615-1_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3613-7

  • Online ISBN: 978-1-4939-3615-1

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