Login Register Cart 0
General Research Article

Characterization of Induced Pluripotent Stem Cells from Human Epidermal Melanocytes by Transduction with Two Combinations of Transcription Factors

Author(s): Sai Cheng, Di Li, Ru-Zhi Zhang*, Jing Zhu, Li Wang, Qi Liu, Ren-He Chen and Xiao-Ming Liu

Volume 19, Issue 6, 2019

Page: [395 - 403] Pages: 9

DOI: 10.2174/1566523220666200211105228

Price: $65

Abstract

Objective: In order to generate induced Pluripotent Stem Cells (iPSCs) more efficiently, it is crucial to identify somatic cells that are easily accessible and possibly require fewer factors for conversion into iPSCs.

Methods: Human epidermal melanocytes were transduced with lentiviral vectors carrying 3 transcription factors (OCT-4, KLF-4 and c-MYC, 3F) or 4 transcription factors (OCT-4, KLF-4, c-MYC and SOX-2, 4F). Once the clones had formed, assays related to stem cell pluripotency, including alkaline phosphatase staining, DNA methylation levels, expression of stem cell markers and ultrastructure analysis were carried out. The iPSCs obtained were then induced to differentiate into the cells representing the three embryonic layers in vitro.

Results: Seven days after the transduction of epidermal melanocytes with 3F or 4F, clones were formed that were positive for alkaline phosphatase staining. Fluorescent staining with antibodies against OCT-4 and SOX-2 was strongly positive, and the cells showed a high nucleus-cytoplasm ratio and active karyokinesis. No melanosomes were found in the cytoplasm by ultrastructural analysis. There were obvious differences in DNA methylation levels between the cloned cells and their parental cells. However, there was not a significant difference between 3F or 4F transfected clonal cells. Meanwhile, the iPSCs successfully differentiated into the three germ layer cells in vitro.

Conclusion: Human epidermal melanocytes do not require ectopic SOX-2 expression for conversion into iPSCs, and may serve as an alternative source for deriving patient-specific iPSCs with fewer genetic elements.

Keywords: Epidermal, melanocytes, multipotential, reprogramming, iPSCS, transcription.

« Previous Next »
Graphical Abstract

[1]
Malik N, Rao MS. A review of the methods for human iPSC derivation. Methods Mol Biol 2013; 997: 23-33.
[http://dx.doi.org/10.1007/978-1-62703-348-0_3] [PMID: 23546745]
[2]
Kawakami T, Okano T, Takeuchi S, et al. Approach for the derivation of melanocytes from induced pluripotent stem cells. J Invest Dermatol 2018; 138(1): 150-8.
[http://dx.doi.org/10.1016/j.jid.2017.07.849] [PMID: 28887108]
[3]
Mortada I, Bilani N. Advances in the production and application of induced pluripotent stem cells. Curr Stem Cell Res Ther 2017; 12(8): 637-43.
[http://dx.doi.org/10.2174/1574888X12666170927115317] [PMID: 28969537]
[4]
Li J, Yin X, Luan Q. Comparative study of periodontal differentiation propensity of induced pluripotent stem cells from different tissue origins. J Periodontol 2018; 89(10): 1230-40.
[http://dx.doi.org/10.1002/JPER.18-0033] [PMID: 30039603]
[5]
Nishihara K, Shiga T, Nakamura E, et al. Induced pluripotent stem cells reprogrammed with three inhibitors show accelerated differentiation potentials with high levels of 2-cell stage marker expression. Stem Cell Reports 2019; 12(2): 305-18.
[http://dx.doi.org/10.1016/j.stemcr.2018.12.018] [PMID: 30713040]
[6]
Wu R, Du D, Bo Y, Zhang M, Zhang L, Yan Y. Hsp90α promotes the migration of iPSCs-derived keratinocyte to accelerate deep second-degree burn wound healing in mice. Biochem Biophys Res Commun 2019; 520(1): 145-51.
[http://dx.doi.org/10.1016/j.bbrc.2019.09.120] [PMID: 31582220]
[7]
Sugiyama-Nakagiri Y, Fujimura T, Moriwaki S. Induction of skin-derived precursor cells from human induced pluripotent stem cells. PLoS One 2016; 11(12): e0168451
[http://dx.doi.org/10.1371/journal.pone.0168451] [PMID: 27992514]
[8]
Utikal J, Maherali N, Kulalert W, Hochedlinger K. Sox2 is dispensable for the reprogramming of melanocytes and melanoma cells into induced pluripotent stem cells. J Cell Sci 2009; 122(Pt 19): 3502-10.
[http://dx.doi.org/10.1242/jcs.054783] [PMID: 19723802]
[9]
Popovic A, Wiggins T, Davids LM. Differential susceptibility of primary cultured human skin cells to hypericin PDT in an in vitro model. J Photochem Photobiol B 2015; 149: 249-56.
[http://dx.doi.org/10.1016/j.jphotobiol.2015.06.009] [PMID: 26114219]
[10]
Adema GJ, de Boer AJ, van ’t Hullenaar R, et al. Melanocyte lineage-specific antigens recognized by monoclonal antibodies NKI-beteb, HMB-50, and HMB-45 are encoded by a single cDNA. Am J Pathol 1993; 143(6): 1579-85.
[PMID: 7504884]
[11]
Raab S, Klingenstein M, Liebau S, Linta L. A comparative view on human somatic cell sources for iPSC generation. Stem Cells Int 2014; 2014: 768391
[http://dx.doi.org/10.1155/2014/768391] [PMID: 25431601]
[12]
Buganim Y, Markoulaki S, van Wietmarschen N, et al. The developmental potential of iPSCs is greatly influenced by reprogramming factor selection. Cell Stem Cell 2014; 15(3): 295-309.
[http://dx.doi.org/10.1016/j.stem.2014.07.003] [PMID: 25192464]
[13]
Streckfuss-Bömeke K, Wolf F, Azizian A, et al. Comparative study of human-induced pluripotent stem cells derived from bone marrow cells, hair keratinocytes, and skin fibroblasts. Eur Heart J 2013; 34(33): 2618-29.
[http://dx.doi.org/10.1093/eurheartj/ehs203] [PMID: 22798560]
[14]
Liu Q, Zhang RZ, Li D, et al. Muse cells, a new type of pluripotent stem cell derived from human fibroblasts. Cell Reprogram 2016; 18(2): 67-77.
[http://dx.doi.org/10.1089/cell.2015.0085] [PMID: 27055628]
[15]
Begemann I, Galic M. Correlative light electron microscopy: Connecting synaptic structure and function. Front Synaptic Neurosci 2016; 8: 28.
[http://dx.doi.org/10.3389/fnsyn.2016.00028] [PMID: 27601992]
[16]
Badja C, Maleeva G, El-Yazidi C, et al. Efficient and cost-effective generation of mature neurons from human induced pluripotent stem cells. Stem Cells Transl Med 2014; 3(12): 1467-72.
[http://dx.doi.org/10.5966/sctm.2014-0024] [PMID: 25355730]
[17]
Muchkaeva IA, Dashinimaev EB, Artyuhov AS, et al. Generation of iPS cells from human hair follice dermal papilla cells. Acta Naturae 2014; 6(1): 45-53.
[http://dx.doi.org/10.32607/20758251-2014-6-1-45-53] [PMID: 24772326]
[18]
Novak D, Weina K, Utikal J. From skin to other cell types of the body. J Dtsch Dermatol Ges 2014; 12(9): 789-92.
[http://dx.doi.org/10.1111/ddg.12403] [PMID: 25087921]
[19]
Khazaei M, Ahuja CS, Fehlings MG. Induced pluripotent stem cells for traumatic spinal cord injury. Front Cell Dev Biol 2017; 4: 152.
[http://dx.doi.org/10.3389/fcell.2016.00152] [PMID: 28154814]
[20]
Abou-Saleh H, Zouein FA, El-Yazbi A, et al. The march of pluripotent stem cells in cardiovascular regenerative medicine. Stem Cell Res Ther 2018; 9(1): 201.
[http://dx.doi.org/10.1186/s13287-018-0947-5] [PMID: 30053890]
[21]
Galach M, Utikal J. From skin to the treatment of diseases--the possibilities of iPS cell research in dermatology. Exp Dermatol 2011; 20(6): 523-8.
[http://dx.doi.org/10.1111/j.1600-0625.2011.01282.x] [PMID: 21585557]
[22]
Larribere L, Utikal J. De- and re-differentiation of the melanocytic lineage. Eur J Cell Biol 2014; 93(1-2): 30-5.
[http://dx.doi.org/10.1016/j.ejcb.2013.11.006] [PMID: 24365127]
[23]
Maherali N, Ahfeldt T, Rigamonti A, Utikal J, Cowan C, Hochedlinger K. A high-efficiency system for the generation and study of human induced pluripotent stem cells. Cell Stem Cell 2008; 3(3): 340-5.
[http://dx.doi.org/10.1016/j.stem.2008.08.003] [PMID: 18786420]
[24]
Saitoh I, Inada E, Iwase Y, et al. Choice of feeders is important when first establishing iPSCs derived from primarily cultured human deciduous tooth dental pulp cells. Cell Med 2015; 8(1-2): 9-23.
[http://dx.doi.org/10.3727/215517915X689038] [PMID: 26858904]
[25]
Eminli S, Utikal J, Arnold K, Jaenisch R, Hochedlinger K. Reprogramming of neural progenitor cells into induced pluripotent stem cells in the absence of exogenous Sox2 expression. Stem Cells 2008; 26(10): 2467-74.
[http://dx.doi.org/10.1634/stemcells.2008-0317] [PMID: 18635867]
[26]
Kar S, Parbin S, Deb M, et al. Epigenetic choreography of stem cells: the DNA demethylation episode of development. Cell Mol Life Sci 2014; 71(6): 1017-32.
[http://dx.doi.org/10.1007/s00018-013-1482-2] [PMID: 24114325]
[27]
Han JW, Yoon YS. Epigenetic landscape of pluripotent stem cells. Antioxid Redox Signal 2012; 17(2): 205-23.
[http://dx.doi.org/10.1089/ars.2011.4375] [PMID: 22044221]
[28]
Takamiya M, Xu F, Suhonen H, et al. Melanosomes in pigmented epithelia maintain eye lens transparency during zebrafish embryonic development. Sci Rep 2016; 6: 25046.
[http://dx.doi.org/10.1038/srep25046] [PMID: 27141993]

Rights & Permissions Print Cite
Article Metrics
14
© 2024 Bentham Science Publishers | Privacy Policy