Quantitative analysis of melanin content in a three-dimensional melanoma cell culture

Chung, Soobin; Lim, Gippeum J.; Lee, Ji Youn

doi:10.1038/s41598-018-37055-y

Cited by 58 publications

(30 citation statements)

References 39 publications

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“…Cloudman S91 is one of the several melanized murine cell lines, with others being B16, B16-F1, B16-F10, and Harding-Passey cells [15][16][17]. The degree of melanization of these cell lines in vitro depends on the presence of several stimulants of melanin synthesis such as alpha-melanocyte stimulating hormone (α-MSH), retinoic acid, etc., [15,16]. To the best of our knowledge, there has not been a quantitative comparison of melanin amount produced by these cell lines when used for tumor initiation in mice.…”

Section: Discussionmentioning

confidence: 99%

Evaluating the Combination of Radioimmunotherapy and Immunotherapy in a Melanoma Mouse Model

Jiao

Allen

Malo

et al. 2020

IJMS

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Immunotherapy has changed the oncology landscape during the last decade and become standard of care for several cancers. The combinations of immunotherapy with other treatment modalities are also being investigated. One of the challenges to investigate such combinations is to identify suitable mouse models for the pre-clinical experiments. In the past, we and other researchers showed that murine B16-F10 melanoma in C57Bl6 mice is refractory to treatment with immune checkpoint inhibitors. In this work we studied the suitability of an alternative syngeneic model, Cloudman S91 murine melanoma in DBA/2 mouse (DBA/2NCrl), to study the combination of immunotherapy targeting PD-1 and radioimmunotherapy targeting melanin. DBA/2 male and female mice were injected subcutaneously with 3–6 million Cloudman S91 cells. When the tumors reached ~150 mm3 volume, the animals were treated intraperitoneally with PBS (sham), h8C3 unlabeled (cold) antibody to melanin, immunotherapy with anti-PD-1 antibody, radioimmunotherapy with 213Bismuth (213Bi)-labeled h8C3 antibody, or several combinations of immunotherapy and radioimmunotherapy. Treatments with immunotherapy alone produced very modest effect on the tumor size, while combination therapy resulted in significant slowing down of the tumor growth, increased animal survival, and no decrease in animal body weight. We conclude that Cloudman S91 murine melanoma in DBA/2 mouse is a suitable model to evaluate combination of immunotherapy of melanoma with tangentially targeted treatments.

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Section: Discussionmentioning

confidence: 99%

Evaluating the Combination of Radioimmunotherapy and Immunotherapy in a Melanoma Mouse Model

Jiao

Allen

Malo

et al. 2020

IJMS

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“…Furthermore, for eventual image-based quantification of cell death in 3D culture using live/dead fluorescent staining, MC38 spheroids were more straightforward than B16-F10 spheroids. As the B16-F10 cells formed spheroids they appeared to produce melanin, an observation recently reported (Chung, Lim, and Lee 2019) . When we stained B16-F10 spheroids in collagen gel with AO/PI live/dead stain, the melanin deposits in the cells masked the fluorescent signal from the stain, making quantification of the fluorescent signals difficult (Supplemental Figure 9).…”

Section: B16-f10 Cells Can Be Replaced With Hgp100-pulsed Mc38 Cellsmentioning

confidence: 64%

Towards a scaled-up T cell-mediated cytotoxicity assay in 3D cell culture using microscopy

Gottschalk

Czech

et al. 2019

Preprint

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Three-dimensional (3D) cell culture systems with tumor spheroids are being adopted for research on the antitumor activity of drug treatments and cytotoxic T cells. Analysis of the cytotoxic effect on 3D tumor cultures within a 3D scaffold, such as collagen, is challenging. Image-based approaches often use confocal microscopy, which greatly limits the sample size of tumor spheroids that can be assayed. We explored a system where tumor spheroids growing in a collagen gel within a microfluidics chip can be treated with drugs or co-cultured with T cells. We attempted to adapt the system to measure the death of cells in the tumor spheroids directly in the microfluidics chip via automated widefield fluorescence microscopy. We were able to successfully measure drug-induced cytotoxicity in tumor spheroids, but had difficulties extending the system to measure T cell-mediated tumor killing.

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“…A comparison to other already developed alpha-MSH peptide analogs is difficult, as for most of them no dose-dependent cellular activity is given [61], or they were tested in melanocytes in vitro [31]. A recent report showed a dose-dependent increase of melanin synthesis in mouse melanoma cells by alpha-MSH reaching a plateau at around 1 to 10 nM [62]. Interestingly, it was found that POMC-derived peptides, such as alpha-MSH and beta-endorphin, could modulate the expression of their respective receptors [45,63].…”

Section: Discussionmentioning

confidence: 99%

Discovery of a Highly Selective MC1R Agonists Pentapeptide to Be Used as a Skin Pigmentation Enhancer and with Potential Anti-Aging Properties

Jackson

Heidl

Imfeld

et al. 2019

IJMS

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One of the first lines of cutaneous defense against photoaging is (a) the synthesis of melanin and (b) the initiation of an oxidative stress response to protect skin against the harmful effects of solar radiation. Safe and selective means to stimulate epidermal pigmentation associated with oxidative stress defense are; however, scarce. Activation of the melanocortin-1 receptor (MC1R) on epidermal melanocytes represents a key step in cutaneous pigmentation initiation and, additionally, it regulates cellular defense mechanisms like oxidative stress and DNA-repair. Thus, making the activation of MC1R an attractive strategy for modulating skin pigmentation and oxidative stress. In this context, we designed and synthesized pentapeptides that act as MC1R agonists. These peptides bound, with high potency, to MC1R and activated cAMP synthesis in CHO cells expressing human MC1R. Using one lead pentapeptide, we could show that this activation of MC1R was specific as testing the activation of other G-protein coupled receptors, including the MC-receptor family, was negative. In vitro efficacy on mouse melanoma cells showed similar potency as for the synthetic MC1R agonist alpha-melanocyte stimulating hormone (NDP-alpha-MSH). Moreover, we could reproduce this activity in human skin tissue culture. The lead pentapeptide was able to induce ex-vivo protein expression of key melanogenesis markers melanocyte inducing transcription factor (MITF), tyrosinase (TYR), and tyrosinase-related protein 1 (TYRP-1). Concerning oxidative stress response, we found that the pentapeptide enhanced the activation of Nrf2 after UVA-irradiation. Our results make this pentapeptide an ideal candidate as a skin pigmentation enhancer that mimics alpha-MSH and may also have anti-photoaging effects on the skin.

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Quantitative analysis of melanin content in a three-dimensional melanoma cell culture

Cited by 58 publications

References 39 publications

Evaluating the Combination of Radioimmunotherapy and Immunotherapy in a Melanoma Mouse Model

Evaluating the Combination of Radioimmunotherapy and Immunotherapy in a Melanoma Mouse Model

Towards a scaled-up T cell-mediated cytotoxicity assay in 3D cell culture using microscopy

Discovery of a Highly Selective MC1R Agonists Pentapeptide to Be Used as a Skin Pigmentation Enhancer and with Potential Anti-Aging Properties

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