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Amir Mishan M, Balagholi S, Chamani T, Feizi S, Soheili Z-S, Rezaei Kanavi M. Potential Effect of Human Platelet Lysate on in vitro Expansion of Human Corneal Endothelial Cells Compared with Y-27632 ROCK Inhibitor. JOVR [Internet]. 2021Jul.29 [cited 2024Apr.18];16(3):349–356. Available from: https://knepublishing.com/index.php/JOVR/article/view/9431
https://doi.org/10.18502/jovr.v16i3.9431
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authors
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Mohammad Amir Mishan
Mohammad Amir Mishan
Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
-
Sahar Balagholi
Sahar Balagholi
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran 3Central Eye Bank of Iran, Tehran, Iran
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Tahereh Chamani
Tahereh Chamani
Central Eye Bank of Iran, Tehran, Iran
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Sepehr Feizi
Sepehr Feizi
Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Zahra-Soheila Soheili
Zahra-Soheila Soheili
National Institute of Genetic Engineering and Biotechnology, Tehran, Iran
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Mozhgan Rezaei Kanavi
Mozhgan Rezaei Kanavi
Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Published Date
- Jul 29, 2021
Potential Effect of Human Platelet Lysate on in vitro Expansion of Human Corneal Endothelial Cells Compared with Y-27632 ROCK Inhibitor
Abstract
Purpose: Corneal endothelial cell (CEC) therapy can be used as a promising therapeutic option for patients with various corneal endothelial dysfunctions. In this study, we compared the proliferative effect of human platelet lysate (HPL), as a xeno-free medium supplement, with Y-27632 Rho/rho-associated protein kinase (ROCK) inhibitor, as a wellknown proliferative and adhesive agent for CECs, and fetal bovine serum (FBS) as the control, in the culture medium of human corneal endothelial cells (HCECs).
Methods: We isolated HCECs from human donors and treated the cells as three different treatment groups including 20% HPL only, 10 μM Y-27632 ROCK inhibitor, combination of 20% HPL and 10 μM Y-27632 ROCK inhibitor, and 20% FBS as the control group. ELISA cell proliferation assay and cell counting was performed on the treated cells. Finally, HCECs were characterized by morphology and immunocytochemistry (ICC).
Results: There was no significant proliferative effect of HPL on cell proliferation compared with the cells treated with Y-27632 ROCK inhibitor or the combination of HPL and Y-27632 ROCK inhibitor, but all the respected treatments had significant inducible effect on cell proliferation as compared with FBS-treated cells. The cells grown in all three treatment groups exhibited CEC morphology. Also, there was a higher expression of Na+/K+-ATPase and ZO-1, as CEC characteristic markers, in the culture of HCECs treated with HPL as compared with FBS.
Conclusion: HPL offers a xeno−free and affordable medium supplement for CEC expansion that can be used in clinical applications.
Keywords:
Cell Proliferation, Corneal Endothelial Cells, Human Platelet Lysate, ROCK Inhibitor
References
1. Dawson DG, Ubels JL, Edelhauser HF. Cornea and sclera. In: Levin LA, Nilsson SFE, Ver Hoeve J, Wu SM, editors. Adler’s physiology of the eye. Elsevier; 2011.
2. Bourne WM. Clinical estimation of corneal endothelial pump function. Trans Am Ophthalmol Soc 1998;96:229– 242.
3. Tan DT, Dart JK, Holland EJ, Kinoshita S. Corneal transplantation. Lancet 2012;379:1749–1761.
4. Gain P, Jullienne R, He Z, Aldossary M, Acquart S, Cognasse F, et al. Global survey of corneal transplantation and eye banking. JAMA Ophthalmol 2016;134:167–173.
5. Parekh M, Ahmad S, Ruzza A, Ferrari S. Human corneal endothelial cell cultivation from old donor corneas with forced attachment. Sci Rep 2017;7:142.
6. Nuzzi R, Marolo P, Tridico F. From DMEK to corneal endothelial cell therapy: technical and biological aspects. J Ophthalmol 2018;2018: 6482095.
7. Kinoshita S, Koizumi N, Ueno M, Okumura N, Imai K, Tanaka H, et al. Injection of cultured cells with a ROCK inhibitor for bullous keratopathy. N Engl J Med 2018;378:995–1003.
8. Okumura N, Koizumi N, Ueno M, Sakamoto Y, Takahashi H, Tsuchiya H, et al. ROCK inhibitor converts corneal endothelial cells into a phenotype capable of regenerating in vivo endothelial tissue. Am J Pathol 2012;181:268–277.
9. Coleman ML, Marshall CJ, Olson MF. RAS and RHO GTPases in G1-phase cell-cycle regulation. Nat Rev Mol Cell Biol 2004;5:355–366.
10. Sun CC, Chiu HT, Lin YF, Lee KY, Pang JHS. Y-27632, a ROCK inhibitor, promoted limbal epithelial cell proliferation and corneal wound healing. PLoS One 2015;10:e0144571.
11. Chapman S, McDermott DH, Shen K, Jang MK, McBride AA. The effect of Rho kinase inhibition on long-term keratinocyte proliferation is rapid and conditional. Stem Cell Res Ther 2014;5:60.
12. Wang T, Kang W, Du L, Ge S. Rho-kinase inhibitor Y-27632 facilitates the proliferation, migration and pluripotency of human periodontal ligament stem cells. J Cell Mol Med 2017;21:3100–3112.
13. Okumura N, Ueno M, Koizumi N, Sakamoto Y, Hirata K, Hamuro J, et al. Enhancement on primate corneal endothelial cell survival in vitro by a ROCK inhibitor. Invest Ophthalmol Vis Sci 2009;50:3680–3687.
14. Liao JK, Seto M, Noma K. Rho kinase (ROCK) inhibitors. J Cardiovasc Pharmacol 2007;50:17–24.
15. Feizi S, Soheili ZS, Bagheri A, Balagholi S, Mohammadian A, Rezaei-Kanavi M, et al. Effect of amniotic fluid on the in vitro culture of human corneal endothelial cells. Exp Eye Res 2014;122:132–140.
16. Thieme D, Reuland L, Lindl T, Kruse F, Fuchsluger T. Optimized human platelet lysate as novel basis for a serum-, xeno-, and additive-free corneal endothelial cell and tissue culture. J Tissue Eng Regen Med 2018;12:557– 564.
17. Okumura N, Sakamoto Y, Fujii K, Kitano J, Nakano S, Tsujimoto Y, et al. Rho kinase inhibitor enables cellbased therapy for corneal endothelial dysfunction. Sci Rep 2016;6:26113.
18. Peh GS, Adnan K, George BL, Ang HP, Seah XY, Tan DT, et al. The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach. Sci Rep 2015;5:9167.
19. Sample SJ, Racette MA, Hans EC, Volstad NJ, Schaefer SL, Bleedorn JA, et al. Use of a platelet-rich plasmacollagen scaffold as a bioenhanced repair treatment for management of partial cruciate ligament rupture in dogs. PloS One 2018;13:e0197204.
20. Spanò R, Muraglia A, Todeschi MR, Nardini M, Strada P, Cancedda R, et al. Platelet-rich plasma-based bioactive membrane as a new advanced wound care tool. J Tissue Eng Regen Med 2018;12:e82–e96.
21. Fernandes G, Yang S. Application of platelet-rich plasma with stem cells in bone and periodontal tissue engineering. Bone Res 2016;4:16036.
22. Naskou MC, Sumner SM, Chocallo A, Kemelmakher H, Thoresen M, Copland I, et al. Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells. Stem Cell Res Ther 2018;9:75.
23. Švajger U. Human platelet lysate is a successful alternative serum supplement for propagation of monocyte-derived dendritic cells. Cytotherapy 2017;19:486–499.
24. Saury C, Lardenois A, Schleder C, Leroux I, Lieubeau B, David L, et al. Human serum and platelet lysate are appropriate xeno-free alternatives for clinical-grade production of human MuStem cell batches. Stem Cell Res Ther 2018;9:128.
25. Chamani T, Javadi MA, Kanavi MR. Trephine-and dyefree technique for eye bank preparation of pre-stripped Descemet membrane endothelial keratoplasty tissue. Cell Tissue Bank 2019;20:321–326.
26. Şeker Ş, Elçin AE, Elçin YM. Autologous protein-based scaffold composed of platelet lysate and aminated hyaluronic acid. J Mater Sci Mater Med 2019;30:127.
27. Su CC, Chen CW, Ho WT, Hu FR, Lee SH, Wang IJ. Phenotypes of trypsin-and collagenase-prepared bovine corneal endothelial cells in the presence of a selective Rho kinase inhibitor, Y-27632. Mol Vis 2015;21:633–643.
28. Pipparelli A, Arsenijevic Y, Thuret G, Gain P, Nicolas M, Majo F. ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells. PloS One 2013;8:e62095.
29. Rasband WS. ImageJ. Bethesda, Maryland, USA: National Institutes of Health; 1997–2016.
30. Strandberg G, Sellberg F, Sommar P, Ronaghi M, Lubenow N, Knutson F, et al, Standardizing the freezethaw preparation of growth factors from platelet lysate. Transfusion 2017;57:1058–1065.
31. Carducci A, Scafetta G, Siciliano C, Carnevale R, Rosa P, Coccia A, et al. GMP-grade platelet lysate enhances proliferation and migration of tenon fibroblasts. Front Biosci 2016;8:84–99.
32. Burnouf T, Strunk D, Koh MB, Schallmoser K. Human platelet lysate: replacing fetal bovine serum as a gold standard for human cell propagation? Biomaterials 2016;76:371–387.
33. Chou ML, Burnouf T, Wang TJ. Ex vivo expansion of bovine corneal endothelial cells in xeno-free medium supplemented with platelet releasate. PloS One 2014;9:e99145.
34. Wang TJ, Chen MS, Chou ML, Lin HC, Seghatchian J, Burnouf T. Comparison of three human platelet lysates used as supplements for in vitro expansion of corneal endothelium cells. Transfus Apher Sci 2017;56:769–773.
35. Olson MF, Ashworth A, Hall A. An essential role for Rho, Rac, and Cdc42 GTPases in cell cycle progression through G1. Science 1995;269:1270–1272.
36. Coleman ML, Olson MF. Rho GTPase signalling pathways in the morphological changes associated with apoptosis. Cell Death Differ 2002;9:493–504.
37. Riento K, Ridley AJ. Rocks: multifunctional kinases in cell behaviour. Nat Rev Mol Cell Biol 2003;4:446–456.
38. Amano M, Fukata Y, Kaibuchi K. Regulation and functions of Rho-associated kinase. Exp Cell Res 2000;261:44–51.
39. Okumura N, Koizumi N, Ueno M, Sakamoto Y, Takahashi H, Hamuro J, et al. The new therapeutic concept of using a rho kinase inhibitor for the treatment of corneal endothelial dysfunction. Cornea 2011;30:S54–S59.
40. Enomoto K, Mimura T, Harris DL, Joyce NC. Age differences in cyclin-dependent kinase inhibitor expression and rb hyperphosphorylation in human corneal endothelial cells. Invest Ophthalmol Vis Sci 2006;47:4330–4340.
41. Sánchez M, Anitua E, Delgado D, Sanchez P, Prado R, Orive G, et al., Platelet-rich plasma, a source of autologous growth factors and biomimetic scaffold for peripheral nerve regeneration. Expert Opin Biol Ther 2017;17:197– 212.
42. Vis PW, Bouten CV, Sluijter JP, Pasterkamp G, van Herwerden LA, Kluin J. Platelet-lysate as an autologous alternative for fetal bovine serum in cardiovascular tissue engineering. Tissue Eng Part A 2010;16:1317–1327.
2. Bourne WM. Clinical estimation of corneal endothelial pump function. Trans Am Ophthalmol Soc 1998;96:229– 242.
3. Tan DT, Dart JK, Holland EJ, Kinoshita S. Corneal transplantation. Lancet 2012;379:1749–1761.
4. Gain P, Jullienne R, He Z, Aldossary M, Acquart S, Cognasse F, et al. Global survey of corneal transplantation and eye banking. JAMA Ophthalmol 2016;134:167–173.
5. Parekh M, Ahmad S, Ruzza A, Ferrari S. Human corneal endothelial cell cultivation from old donor corneas with forced attachment. Sci Rep 2017;7:142.
6. Nuzzi R, Marolo P, Tridico F. From DMEK to corneal endothelial cell therapy: technical and biological aspects. J Ophthalmol 2018;2018: 6482095.
7. Kinoshita S, Koizumi N, Ueno M, Okumura N, Imai K, Tanaka H, et al. Injection of cultured cells with a ROCK inhibitor for bullous keratopathy. N Engl J Med 2018;378:995–1003.
8. Okumura N, Koizumi N, Ueno M, Sakamoto Y, Takahashi H, Tsuchiya H, et al. ROCK inhibitor converts corneal endothelial cells into a phenotype capable of regenerating in vivo endothelial tissue. Am J Pathol 2012;181:268–277.
9. Coleman ML, Marshall CJ, Olson MF. RAS and RHO GTPases in G1-phase cell-cycle regulation. Nat Rev Mol Cell Biol 2004;5:355–366.
10. Sun CC, Chiu HT, Lin YF, Lee KY, Pang JHS. Y-27632, a ROCK inhibitor, promoted limbal epithelial cell proliferation and corneal wound healing. PLoS One 2015;10:e0144571.
11. Chapman S, McDermott DH, Shen K, Jang MK, McBride AA. The effect of Rho kinase inhibition on long-term keratinocyte proliferation is rapid and conditional. Stem Cell Res Ther 2014;5:60.
12. Wang T, Kang W, Du L, Ge S. Rho-kinase inhibitor Y-27632 facilitates the proliferation, migration and pluripotency of human periodontal ligament stem cells. J Cell Mol Med 2017;21:3100–3112.
13. Okumura N, Ueno M, Koizumi N, Sakamoto Y, Hirata K, Hamuro J, et al. Enhancement on primate corneal endothelial cell survival in vitro by a ROCK inhibitor. Invest Ophthalmol Vis Sci 2009;50:3680–3687.
14. Liao JK, Seto M, Noma K. Rho kinase (ROCK) inhibitors. J Cardiovasc Pharmacol 2007;50:17–24.
15. Feizi S, Soheili ZS, Bagheri A, Balagholi S, Mohammadian A, Rezaei-Kanavi M, et al. Effect of amniotic fluid on the in vitro culture of human corneal endothelial cells. Exp Eye Res 2014;122:132–140.
16. Thieme D, Reuland L, Lindl T, Kruse F, Fuchsluger T. Optimized human platelet lysate as novel basis for a serum-, xeno-, and additive-free corneal endothelial cell and tissue culture. J Tissue Eng Regen Med 2018;12:557– 564.
17. Okumura N, Sakamoto Y, Fujii K, Kitano J, Nakano S, Tsujimoto Y, et al. Rho kinase inhibitor enables cellbased therapy for corneal endothelial dysfunction. Sci Rep 2016;6:26113.
18. Peh GS, Adnan K, George BL, Ang HP, Seah XY, Tan DT, et al. The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach. Sci Rep 2015;5:9167.
19. Sample SJ, Racette MA, Hans EC, Volstad NJ, Schaefer SL, Bleedorn JA, et al. Use of a platelet-rich plasmacollagen scaffold as a bioenhanced repair treatment for management of partial cruciate ligament rupture in dogs. PloS One 2018;13:e0197204.
20. Spanò R, Muraglia A, Todeschi MR, Nardini M, Strada P, Cancedda R, et al. Platelet-rich plasma-based bioactive membrane as a new advanced wound care tool. J Tissue Eng Regen Med 2018;12:e82–e96.
21. Fernandes G, Yang S. Application of platelet-rich plasma with stem cells in bone and periodontal tissue engineering. Bone Res 2016;4:16036.
22. Naskou MC, Sumner SM, Chocallo A, Kemelmakher H, Thoresen M, Copland I, et al. Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells. Stem Cell Res Ther 2018;9:75.
23. Švajger U. Human platelet lysate is a successful alternative serum supplement for propagation of monocyte-derived dendritic cells. Cytotherapy 2017;19:486–499.
24. Saury C, Lardenois A, Schleder C, Leroux I, Lieubeau B, David L, et al. Human serum and platelet lysate are appropriate xeno-free alternatives for clinical-grade production of human MuStem cell batches. Stem Cell Res Ther 2018;9:128.
25. Chamani T, Javadi MA, Kanavi MR. Trephine-and dyefree technique for eye bank preparation of pre-stripped Descemet membrane endothelial keratoplasty tissue. Cell Tissue Bank 2019;20:321–326.
26. Şeker Ş, Elçin AE, Elçin YM. Autologous protein-based scaffold composed of platelet lysate and aminated hyaluronic acid. J Mater Sci Mater Med 2019;30:127.
27. Su CC, Chen CW, Ho WT, Hu FR, Lee SH, Wang IJ. Phenotypes of trypsin-and collagenase-prepared bovine corneal endothelial cells in the presence of a selective Rho kinase inhibitor, Y-27632. Mol Vis 2015;21:633–643.
28. Pipparelli A, Arsenijevic Y, Thuret G, Gain P, Nicolas M, Majo F. ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells. PloS One 2013;8:e62095.
29. Rasband WS. ImageJ. Bethesda, Maryland, USA: National Institutes of Health; 1997–2016.
30. Strandberg G, Sellberg F, Sommar P, Ronaghi M, Lubenow N, Knutson F, et al, Standardizing the freezethaw preparation of growth factors from platelet lysate. Transfusion 2017;57:1058–1065.
31. Carducci A, Scafetta G, Siciliano C, Carnevale R, Rosa P, Coccia A, et al. GMP-grade platelet lysate enhances proliferation and migration of tenon fibroblasts. Front Biosci 2016;8:84–99.
32. Burnouf T, Strunk D, Koh MB, Schallmoser K. Human platelet lysate: replacing fetal bovine serum as a gold standard for human cell propagation? Biomaterials 2016;76:371–387.
33. Chou ML, Burnouf T, Wang TJ. Ex vivo expansion of bovine corneal endothelial cells in xeno-free medium supplemented with platelet releasate. PloS One 2014;9:e99145.
34. Wang TJ, Chen MS, Chou ML, Lin HC, Seghatchian J, Burnouf T. Comparison of three human platelet lysates used as supplements for in vitro expansion of corneal endothelium cells. Transfus Apher Sci 2017;56:769–773.
35. Olson MF, Ashworth A, Hall A. An essential role for Rho, Rac, and Cdc42 GTPases in cell cycle progression through G1. Science 1995;269:1270–1272.
36. Coleman ML, Olson MF. Rho GTPase signalling pathways in the morphological changes associated with apoptosis. Cell Death Differ 2002;9:493–504.
37. Riento K, Ridley AJ. Rocks: multifunctional kinases in cell behaviour. Nat Rev Mol Cell Biol 2003;4:446–456.
38. Amano M, Fukata Y, Kaibuchi K. Regulation and functions of Rho-associated kinase. Exp Cell Res 2000;261:44–51.
39. Okumura N, Koizumi N, Ueno M, Sakamoto Y, Takahashi H, Hamuro J, et al. The new therapeutic concept of using a rho kinase inhibitor for the treatment of corneal endothelial dysfunction. Cornea 2011;30:S54–S59.
40. Enomoto K, Mimura T, Harris DL, Joyce NC. Age differences in cyclin-dependent kinase inhibitor expression and rb hyperphosphorylation in human corneal endothelial cells. Invest Ophthalmol Vis Sci 2006;47:4330–4340.
41. Sánchez M, Anitua E, Delgado D, Sanchez P, Prado R, Orive G, et al., Platelet-rich plasma, a source of autologous growth factors and biomimetic scaffold for peripheral nerve regeneration. Expert Opin Biol Ther 2017;17:197– 212.
42. Vis PW, Bouten CV, Sluijter JP, Pasterkamp G, van Herwerden LA, Kluin J. Platelet-lysate as an autologous alternative for fetal bovine serum in cardiovascular tissue engineering. Tissue Eng Part A 2010;16:1317–1327.