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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
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