Skip to main content

Study of Mesenchymal Stem Cell-Mediated Mitochondrial Transfer in In Vitro Models of Oxidant-Mediated Airway Epithelial and Smooth Muscle Cell Injury

  • Protocol
  • First Online:
In Vitro Models for Stem Cell Therapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2269))

Abstract

Mesenchymal stem cells (MSCs) have emerged as an attractive candidate for cell-based therapy. In the past decade, many animal and pilot clinical studies have demonstrated that MSCs are therapeutically beneficial for the treatment of obstructive lung diseases such as asthma and chronic obstructive pulmonary disease (COPD). However, due to the scarcity of adult human MSCs, human-induced pluripotent stem cells mesenchymal stem cells (iPSCs) are now increasingly used as a source of MSCs. iPSCs are derived by reprogramming somatic cells from a wide variety of tissues such as skin biopsies and then differentiating them into iPSC-MSCs. One of the mechanisms through which MSCs exert their protective effects is mitochondrial transfer. Specifically, transfer of mitochondria from iPSC-MSCs to lung cells was shown to protect lung cells against oxidative stress-induced mitochondrial dysfunction and apoptosis and to reduce lung injury and inflammation in in vivo models of lung disease. In this chapter, we detail our methods to visualize and quantify iPSC-MSC-mediated mitochondrial transfer and to study its effects on oxidant-induced airway epithelial and smooth muscle cell models of acute airway cell injury.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 179.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Wecht S, Rojas M (2016) Mesenchymal stem cells in the treatment of chronic lung disease. Respirology 21(8):1366–1375. https://doi.org/10.1111/resp.12911

    Article  PubMed  Google Scholar 

  2. Harrell CR, Sadikot R, Pascual J et al (2019) Mesenchymal stem cell-based therapy of inflammatory lung diseases: current understanding and future perspectives. Stem Cells Int 2019:4236973. https://doi.org/10.1155/2019/4236973

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Fan XL, Zhang Z, Ma CY et al (2019) Mesenchymal stem cells for inflammatory airway disorders: promises and challenges. Biosci Rep 39(1). https://doi.org/10.1042/BSR20182160

  4. Antunes MA, Lapa ESJR, Rocco PR (2017) Mesenchymal stromal cell therapy in COPD: from bench to bedside. Int J Chron Obstruct Pulmon Dis 12:3017–3027. https://doi.org/10.2147/COPD.S146671

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Broekman W, Khedoe P, Schepers K et al (2018) Mesenchymal stromal cells: a novel therapy for the treatment of chronic obstructive pulmonary disease? Thorax 73(6):565–574. https://doi.org/10.1136/thoraxjnl-2017-210672

    Article  PubMed  Google Scholar 

  6. Kokturk N, Yildirim F, Gulhan PY et al (2018) Stem cell therapy in chronic obstructive pulmonary disease. How far is it to the clinic? Am J Stem Cells 7(3):56–71

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Sun Z, Li F, Zhou X et al (2018) Stem cell therapies for chronic obstructive pulmonary disease: current status of pre-clinical studies and clinical trials. J Thorac Dis 10(2):1084–1098. https://doi.org/10.21037/jtd.2018.01.46

    Article  PubMed  PubMed Central  Google Scholar 

  8. Toraldo DM, Toraldo S, Conte L (2018) The clinical use of stem cell research in chronic obstructive pulmonary disease: a critical analysis of current policies. J Clin Med Res 10(9):671–678. https://doi.org/10.14740/jocmr3484w

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Uccelli A, Moretta L, Pistoia V (2008) Mesenchymal stem cells in health and disease. Nat Rev Immunol 8(9):726–736. https://doi.org/10.1038/nri2395

    Article  CAS  PubMed  Google Scholar 

  10. Li O, Tormin A, Sundberg B et al (2013) Human embryonic stem cell-derived mesenchymal stroma cells (hES-MSCs) engraft in vivo and support hematopoiesis without suppressing immune function: implications for off-the shelf ES-MSC therapies. PLoS One 8(1):e55319. https://doi.org/10.1371/journal.pone.0055319

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Gharibi B, Farzadi S, Ghuman M et al (2014) Inhibition of Akt/mTOR attenuates age-related changes in mesenchymal stem cells. Stem Cells 32(8):2256–2266. https://doi.org/10.1002/stem.1709

    Article  CAS  PubMed  Google Scholar 

  12. Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126(4):663–676. https://doi.org/10.1016/j.cell.2006.07.024

    Article  CAS  PubMed  Google Scholar 

  13. Yu J, Vodyanik MA, Smuga-Otto K et al (2007) Induced pluripotent stem cell lines derived from human somatic cells. Science 318(5858):1917–1920. https://doi.org/10.1126/science.1151526

    Article  CAS  PubMed  Google Scholar 

  14. Dominici M, Le Blanc K, Mueller I et al (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8(4):315–317. https://doi.org/10.1080/14653240600855905

    Article  CAS  PubMed  Google Scholar 

  15. Lian QZ, Zhang YL, Zhang JQ et al (2010) Functional mesenchymal stem cells derived from human induced pluripotent stem cells attenuate limb ischemia in mice. Circulation 121(9):1113–U1191. https://doi.org/10.1161/Circulationaha.109.898312

    Article  PubMed  Google Scholar 

  16. Spees JL, Olson SD, Whitney MJ et al (2006) Mitochondrial transfer between cells can rescue aerobic respiration. Proc Natl Acad Sci U S A 103(5):1283–1288. https://doi.org/10.1073/pnas.0510511103

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Li X, Zhang Y, Yeung SC et al (2014) Mitochondrial transfer of induced pluripotent stem cell-derived mesenchymal stem cells to airway epithelial cells attenuates cigarette smoke-induced damage. Am J Respir Cell Mol Biol 51(3):455–465. https://doi.org/10.1165/rcmb.2013-0529OC

    Article  CAS  PubMed  Google Scholar 

  18. Yao Y, Fan XL, Jiang D et al (2018) Connexin 43-mediated mitochondrial transfer of iPSC-MSCs alleviates asthma inflammation. Stem Cell Rep 11(5):1120–1135. https://doi.org/10.1016/j.stemcr.2018.09.012

    Article  CAS  Google Scholar 

  19. Paliwal S, Chaudhuri R, Agrawal A et al (2018) Regenerative abilities of mesenchymal stem cells through mitochondrial transfer. J Biomed Sci 25(1):31. https://doi.org/10.1186/s12929-018-0429-1

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Li X, Michaeloudes C, Zhang Y et al (2018) Mesenchymal stem cells alleviate oxidative stress-induced mitochondrial dysfunction in the airways. J Allergy Clin Immunol 141(5):1634–1645. e1635. https://doi.org/10.1016/j.jaci.2017.08.017

    Article  CAS  PubMed  Google Scholar 

  21. Lian Q, Zhang Y, Liang X et al (2016) Directed differentiation of human-induced pluripotent stem cells to mesenchymal stem cells. Methods Mol Biol 1416:289–298. https://doi.org/10.1007/978-1-4939-3584-0_17

    Article  CAS  PubMed  Google Scholar 

  22. Michaeloudes C, Chang PJ, Petrou M et al (2011) Transforming growth factor-beta and nuclear factor E2-related factor 2 regulate antioxidant responses in airway smooth muscle cells: role in asthma. Am J Respir Crit Care Med 184(8):894–903

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Michaeloudes C, Kuo CH, Haji G et al (2017) Metabolic re-patterning in COPD airway smooth muscle cells. Eur Respir J 50(5). https://doi.org/10.1183/13993003.00202-2017

  24. Yang NC, Ho WM, Chen YH et al (2002) A convenient one-step extraction of cellular ATP using boiling water for the luciferin-luciferase assay of ATP. Anal Biochem 306(2):323–327. https://doi.org/10.1006/abio.2002.5698

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by a National Natural Science Grant of China (No 81370140 to JCW Mak), the Hong Kong Research Grant Council General Research Fund (HKU 773612M to JCW Mak), and the Sanming Project of Medicine in Shenzhen (SZSM201612096).

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Judith C. W. Mak or Pankaj K. Bhavsar .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Michaeloudes, C., Li, X., Mak, J.C.W., Bhavsar, P.K. (2021). Study of Mesenchymal Stem Cell-Mediated Mitochondrial Transfer in In Vitro Models of Oxidant-Mediated Airway Epithelial and Smooth Muscle Cell Injury. In: Stock, P., Christ, B. (eds) In Vitro Models for Stem Cell Therapy. Methods in Molecular Biology, vol 2269. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1225-5_7

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1225-5_7

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1224-8

  • Online ISBN: 978-1-0716-1225-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics