UV-Irradiated RPE Cells Assist Differentiation of Bone Marrow Derived Mesenchymal Stem Cells into RPE Cells Under a Direct Co-Culture Environment

Min Joung Choi; Gi Won Lee; Jin Su Kim; Han Sol Kim; David Kim; Jeong Eun Song; Muthukumar Thangavelu; Gilson Khang

Macromolecular Research, Vol.27, No.8, 781-788, August, 2019

DOI10.1007/s13233-019-7114-4 Export Citation

UV-Irradiated RPE Cells Assist Differentiation of Bone Marrow Derived Mesenchymal Stem Cells into RPE Cells Under a Direct Co-Culture Environment

Min Joung Choi, Gi Won Lee, Jin Su Kim, Han Sol Kim, David Kim, Jeong Eun Song , Muthukumar Thangavelu, and Gilson Khang^†

Department of BIN Convergence Technology, Department of Polymer Nano Science & Technology and Polymer Materials Fusion Research Center, Chonbuk National University, Deokjin, Jeonju, Jeonbuk 54896, Korea

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Degeneration of retinal pigment epithelium cells (RPE) or delamination of RPE from Bruch’s membrane causes visual impairment. While regeneration of RPE cells is desirable to treat age-related macular degeneration (AMD) RPE cells of mammals, which are inherently non-renewable. Here, the bone marrow derived mesenchymal stem cells (BMSCs) that have the advantage of rapid growth and ease of extraction, which are adopted in order to help regenerate RPE cell. By co-culture with RPE cell, BMSCs exhibited a tendency to differentiate into RPE cell. By irradiating with the ultraviolet (UV) light, RPE cell was inhibited from growing and the differentiation of BMSCs was induced. The B-27® supplement was used as a differentiation factor, which helps BMSCs to differentiation into RPE cells. Specifically, B-27® supplement was mixed into the growth medium, which was added only to the co-culture and BMSCs groups. In consequence, UV-irradiated RPE cells did not grow, BMSCs in the co-culture group exhibited a tendency to differentiate into RPE cells. This was demonstrated by MTT assay, SEM, confocal imaging of the live/dead cells, RT-PCR and histological staining. Cell proliferation was the highest in the co-culture group, and the differentiation of BMSCs into RPE cells was observed in RT-PCR and histological staining. Our method will allow BMSCs with B-27® supplement to offer a new direction for RPE regeneration.

Keywords:RPE;co-culture;BMSC;differentiation;and Gellan-gum

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[Related Articles]

[1] Friedman DS, O'Colmain BJ, Munoz B, Tomany SC, McCarty C, de Jong PT, Nemesure B, Mitchell P, Kempen J, Diseases GE, Prevalence Research, Arch Ophthalmol 122, 564 (2004).

[2] Nazari H, Zhang L, Zhu D, Chader GJ, Falabella P, Stefanini F, Rowland T, Clegg DO, Kashani AH, Hinton DR, Humayun MS, Prog. Retin. Eye. Res., 48, 1 (2015)

[3] Sanchez-Ramos J, Song S, Cardozo-Pelaez F, Hazzi C, Stedeford T, Willing A, Freeman TB, Saporta S, Janssen W, Patel N, Cooper DR, Sanberg PR, Exp. Neurol., 164, 247 (2000)

[4] Duan P, Xu HW, Zeng YX, Wang Y, Yin ZQ, Cell. Physiol. Biochem., 31, 601 (2013)

[5] Oliveira JT, Martins L, Picciochi R, Malafaya PB, Sousa RA, Neves NM, Mano JF, Reis RL, J. Biomed. Mater. Res. A, 93, 852 (2010)

[6] Silva-Correia J, Zavan B, Vindigni V, Silva TH, Oliveira JM, Abatangelo G, Reis RL, Adv. Healthc. Mater., 2, 568 (2013)

[7] Khang G, Lee SK, Kim HN, Silva-Correia J, Gomes ME, Viegas CA, Dias IR, Oliveira JM, Reis RL, J. Tissue. Eng. Regen. Med., 9, 265 (2015)

[8] Vashist A, Vashist A, Gupta YK, Ahmad S, J. Mater. Chem. B., 2, 147 (2014)

[9] Silva-Correia J, Oliveira JM, Caridade SG, Oliveira JT, Sousa RA, Mano JF, Reis RL, J. Tissue. Eng. Regen. Med., 5, e97 (2011)

[10] Behravesh E, Zygourakis K, Mikos AG, J. Biomed. Mater. Res. A, 65, 260 (2013)

[11] Fang H, Song P, Shen Y, Shen C, Liu X, J. Spinal. Cord. Med., 41, 10 (2018)

[12] Wankun X, Wenzhen Y, Min Z, Weiyan Z, Huan C, Wei D, Lvzhen H, Xu Y, Xiaoxin L, Mol. Vis., 17, 3512 (2011)

[13] Dai L, Liu XX, Tong Z, Carbohydr. Polym., 81, 207 (2010)

[14] Ahuja M, Singh S, Kumar A, Int. J. Biological Macromol., 53, 114 (2013)

[15] Causa F, Netti PA, Ambrosio L, Biomaterials, 28, 5093 (2007)

[16] Salerno A, Di Maio E, Iannace S, Netti PA, J. Porous. Mat., 19, 181 (2012)

[17] Tratsk KS, Thanos S, Graefes Arch. Clin. Exp. Ophthalmol., 241, 852 (2003)

[18] Spiller KL, Liu Y, Holloway JL, Maher SA, Cao YL, Liu W, Zhou GD, Lowman AM, J. Control. Release, 157, 39 (2012)

[19] Barnstable CJ, Tombran-Tink J, Prog. Retin. Eye. Res., 23, 561 (2004)

[20] Wu S, Xu R, Duan B, Jiang P, J. Mater. Chem. B, 5, 3870 (2017)

[21] Yang LL, Zhou QJ, Wang Y, Wang YQ, Int. J. Ophthalmol-Chi., 3, 23 (2010)

[22] Chiou SH, Kao CL, Peng CH, Chen SJ, Tarng YW, Ku HH, Chen YC, Shyr YM, Liu RS, Hsu CJ, Yang DM, Hsu WM, Kuo CD, Lee CH, Biochem. Biophys. Res. Commun., 326(3), 578 (2005)

[23] Pfaffl MW, Horgan GW, Dempfle L, Nucleic. Acids. Res., 30, #36 (2002)

[24] Dheda K, Huggett JF, Bustin SA, Johnson MA, Rook G, Zumla A, Biotechniques, 37, 112 (2004)

[25] Park JH, Shin EY, Shin ME, Choi MJ, Carlomagno C, Song JE, Khang G, Int. J. Biol. Macromol., 117, 546 (2018)

[26] Yang F, Williams CG, Wang DA, Lee H, Manson PN, Elisseeff J, Biomaterials, 26, 5991 (2005)

[27] Oliveira JT, Santos TC, Martins L, Picciochi R, Marques AP, Castro AG, Neves NM, Mano JF, Reis RL, Tissue. Eng. Part A., 16, 343 (2010)

[28] Pereira DR, Silva-Correia J, Oliveira JM, Joao J, Helder H, Reis RL, J. Tissue. Eng. Regen. M., 6, 185 (2012)

[29] Canadas R, Pereira DR, Silva-Correia J, Marques AP, Oliveira JM, Reis RL, J. Tissue. Eng. Regen. M., 6, 16 (2012)