Research ArticleExperimental Studies
Open Access

Polysaccharides Derived from Caulerpa lentillifera (Sea Grape) Inhibit Melanogenesis in Human Melanoma Cells and Promote Stemness in Stem Cells

RATANA CHAIKLAHAN, ZIN ZIN EI, NATTAYAPORN CHIRASUWAN, THANYALAK SRINORASING, BHURICHAYA INNETS and PITHI CHANVORACHOTE
In Vivo November 2025, 39 (6) 3258-3270; DOI: https://doi.org/10.21873/invivo.14125

Abstract

Background/Aim: Aging results in diminished physiological functions and reduced stem cell activity, driving research into cellular mechanisms and natural compounds that support tissue regeneration and improve skin health. Marine algae, particularly Caulerpa lentillifera (sea grape), are rich in sulfated polysaccharides and widely used in the food and cosmetics industries due to their diverse biological properties, including UV protection, anti-inflammatory effects, and hydration.

Materials and Methods: The expression levels of stem cell transcription factors were evaluated at the mRNA level using RT-qPCR and at the protein level via western blot and immunofluorescence. Melanin content was quantified in human melanoma cells treated with sea grape extract.

Results: The findings revealed that treatment with sea grape significantly enhanced the expression of stem cell transcription factors in human Dermal Papilla (DP) cells, with mRNA levels of OCT4, NANOG, and SOX2 increasing by 20-, 35-, and 15-fold, respectively. Likewise, in UE6E7T-3 bone marrow stem cells, sea grape induced OCT4, NANOG, and SOX2 induction by 1.5-, 1.5-, and 2.5-fold, respectively. Corresponding protein levels of these stem cell markers also showed increased expression in both sea grape-treated DP cells and UE6E7T-3 cells. Additionally, sea grape exhibited a significant anti-melanogenic effect on human melanoma cells.

Conclusion: Overall, sea grape reduces melanin levels and stimulates regenerative markers in stem cells, presenting a promising opportunity for developing novel products focused on pigmentation regulation and skin rejuvenation.

Keywords:

Introduction

Skin health is profoundly affected by the aging process, which is commonly characterized by hyperpigmentation, hair thinning, and a diminished regenerative capacity (1, 2). Consequently, considerable research efforts are focused on developing strategies that modulate key cellular pathways involved in aging, with the goal of enhancing skin rejuvenation, maintaining pigmentation homeostasis, and preserving hair follicle integrity (3, 4). Skin color is influenced by both intrinsic factors, such as skin type and genetic makeup, and extrinsic factors, including sunlight exposure and environmental pollution (5, 6). Melanogenesis is the process by which melanin is produced, and it is the main factor responsible for human skin pigmentation (7). An overproduction of melanin can lead to various aesthetic issues, including melasma, freckles, and post-inflammatory hyperpigmentation (8).

Previous studies have shown that conditioned medium (CM) derived from stem cells contains various secreted factors, including cytokines, chemokines, growth factors, and extracellular vesicles such as exosomes (9, 10). These secreted factors have been shown to suppress melanin production both directly and via paracrine effects on the skin microenvironment, highlighting the potential of neural stem cell CM as a novel depigmenting agent (9-11).

Stem cells maintain tissue function through self-renewal and differentiation. Aging impairs these capacities, reducing regeneration and implicating stem cell decline in aging (12). However, the contribution of stem cell dysfunction versus systemic tissue degeneration likely varies across tissues (12). As a result, many studies have investigated natural products that enhance stemness by promoting self-renewal and differentiation.

Interest in marine bioactive compounds has grown for natural dermatological and cosmetic uses (13). Marine algae provide sulfated polysaccharides used as gelling agents, thickeners, and emulsifiers in food (14), or in cosmetics for skin moisturizing, anti-aging, and skin tightening (15). Polysaccharide extracts from the brown seaweed Sargassum vachellianum protect skin from UV damage, inhibits tyrosinase, and retain moisture (16). Similarly, polysaccharides from the green seaweed Caulerpa microphysa exhibit anti-inflammatory, wound-healing, and moisturizing effects (17). Awanthi et al. (18) reported that polysaccharides from C. lentillifera inhibit hyaluronidase activity. Among these marine resources, C. lentillifera (sea grape) is notable for its antioxidant, anti-inflammatory, and antimicrobial properties (19) and a higher polysaccharide content than other species (20). Polysaccharide extracts from C. lentillifera have shown significant antioxidant potential in in vitro assays and cellular antioxidant activity studies (21).

However, the uses of polysaccharides from C. lentillifera in skin health and regeneration remain unclear. This study analyzes their biochemical properties and evaluates their effects on melanin regulation and the potential to induce stem cell property in human cells.

Materials and Methods

Preparation of polysaccharide extract. Fresh C. lentillifera (25 kg wet weight, ~1.25 kg dry weight) was sourced from a Caulerpa farm in Samut Sakhon, Thailand. After thorough washing, the algae were homogenized and extracted with 25 l of hot water (1:1 w/v) at 90°C for 60 min (pH ~6.0-7.0). The mixture was centrifuged at 10,000×g, 25°C for 10 min to remove insoluble material. The supernatant containing water-soluble polysaccharides was concentrated by rotary evaporation at 50°C under reduced pressure. The concentrated extract (10-15 mg/ml solids) was spray-dried at 150°C inlet and 75°C outlet temperatures to obtain polysaccharide powder for further analysis (Figure 1). The extraction yield was calculated as the percentage of polysaccharide dry weight relative to the dry weight of the initial algal sample using the following equation: Yield (% of the dry weight)=[weight of polysaccharides (g)] / [weight of sample (g)]×100.

Figure 1:
Figure 1:

Schematic diagram of the preparation of polysaccharide powder from C. lentillifera. The process includes hot water extraction (90°C, 60 min), centrifugation (10,000×g, 25°C, 10 min), vacuum concentration (50°C bath temperature, 2 mbar, 55 rpm), and spray drying (inlet 150°C, outlet 75°C) to obtain a dry polysaccharide powder.

Chemical composition analysis. The spray-dried polysaccharide powder’s composition was analyzed using standard methods (22). Total sugars were measured by the phenol-sulfuric acid assay at 490 nm with glucose as standard. Monosaccharides were identified by HPLC after hydrolysis with trifluoroacetic acid. Protein content was determined using a 2-D Quant Kit with absorbance at 480 nm.

Sulfate content was measured by the barium chloride-gelatin method after acid hydrolysis, with absorbance at 360 nm. Uronic acid was quantified by the m-hydroxydiphenyl method, measuring absorbance at 525 nm. Total phenolics were assessed by Folin-Ciocalteu assay at 765 nm and expressed as mg gallic acid equivalents per gram dry weight.

Cell culture. Primary human dermal papilla (DP) cells, UE6E7T-3 mesenchymal stem cells, and G361 melanoma cells were obtained from commercial sources and cultured in appropriate media: high-glucose DMEM (cat. no. 12800-058, Gibco, Gaithersburg, MD, USA) for DP cells, low-glucose DMEM (cat. no. 31600-026, Gibco) for UE6E7T-3, and McCoy’s 5a medium (cat. no. 30-2007, ATCC, Manassas, VA, USA) for G361 cells. All media were supplemented with 10% fetal bovine serum, 2 mM glutamine, and 1% Antibiotics-Antimycotic (Gibco). Cells were maintained at 37°C in a humidified atmosphere containing 5% CO2. Experimental procedures were conducted when cells reached 70-80% confluence.

Cells and reagents. The reagents 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), bovine serum albumin (BSA), Hoechst 33342, and synthetic melanin (cat no. M8631) were procured from Sigma-Aldrich, Co. (St. Louis, MO, USA). Mouse monoclonal primary antibodies for CD44 (cat. no. 3570) were obtained from Cell Signaling Technology (Beverly, MA, USA). Rabbit monoclonal primary antibodies targeting OCT4 (cat. no. ab19857), NANOG (cat. no. ab80892), SOX2 (cat. no. ab97959), and CD133 (cat. no. ab19898) were purchased from Abcam (Waltham, MA, USA).

Secondary antibodies, including Alexa Fluor™ 594 goat anti-rabbit IgG (H+L) (Cat no: A11037) and Alexa Fluor™ 488 goat anti-mouse IgG (H+L) (Cat no: A11029), were supplied by Invitrogen (Carlsbad, CA, USA). Primers for OCT4, NANOG, SOX2, and GAPDH were sourced from Eurofins Genomics (Louisville, KY, USA).

Cell viability. Cell viability was assessed using the MTT assay. Cells (1×104 cells/well) were seeded in 96-well plates, incubated for 24 h, then treated with sea grape extract (0-200 μg/ml) for another 24 h. After treatment, MTT reagent (4 mg/ml in PBS) was added and incubated for 3 h. Formazan crystals were dissolved in DMSO, and absorbance was measured at 570 nm using a microplate reader.

Quantitative analysis for real-time PCR. Total RNA was extracted from sea grape-treated cells using GENEzol and converted to cDNA using SuperScript III. RT-qPCR was performed with 100 ng cDNA and Luna® Universal qPCR Master Mix on a Bio-Rad CFX 96 system. The protocol included denaturation at 95°C for 1 min, followed by 45 cycles of 95°C for 15 s and 60°C for 30s. Melting curve analysis confirmed primer specificity. Gene expression was normalized to GAPDH and calculated using the comparative Cq method.

Immunofluorescence. Cells (1×104 cell/well) were treated with sea grape extract (0, 25, 50 μg/ml) for 24 h, fixed, permeabilized, and blocked to prevent non-specific binding. They were incubated overnight at 4°C with primary antibodies (1:400), followed by Alexa Fluor-conjugated secondary antibodies and Hoechst 33342 for nuclear staining. After washing and mounting, immunofluorescence images were captured using an Olympus IX51 microscope, and fluorescence intensity was analyzed with ImageJ.

Western blot analysis. Western blotting was used to assess stemness protein expression in cells treated with sea grape extract (0, 25, 50 μg/ml) for 24 h. Cells were lysed in buffer with protease inhibitors, and protein concentration was measured via BCA assay. Proteins were separated by SDS-PAGE, transferred to nitrocellulose membranes, and blocked with 5% milk. Membranes were incubated overnight with primary antibodies against CD133, CD44, OCT4, and SOX2 (β-actin as control), followed by HRP-conjugated secondary antibodies. Detection was performed using chemiluminescent substrate and visualized with the iBright CL1500 system. Band intensities were analyzed using ImageJ.

Determination of melanin content in G361 melanoma cells. G361 melanoma cells (1×105 cell/well) were treated with sea grape extract (0, 10, 20 μg/ml) for 24, 48, and 72 h. After treatment, cells were collected, lysed in 1 N NaOH with 10% DMSO at 80°C for 3 h, and melanin content was measured by absorbance at 405 nm using a microplate reader.

Cellular tyrosinase assay. Tyrosinase activity was measured in G361 melanoma cells treated with sea grape extract (0, 10, 20 μg/ml) for 24, 48, and 72 h. After treatment, cells were lysed with 1% Triton X-100 containing protease inhibitors. Lysates were clarified by centrifugation, and protein concentration was determined via BCA assay. Equal protein amounts were incubated with 2 mM L-DOPA at 37°C for 2 h, and dopachrome formation was measured at 490 nm to assess tyrosinase activity.

Statistical analysis. The data are expressed as the mean ± standard deviation (SD) from three or more independent biological experiments. Multiple comparisons were made using one-way ANOVA, followed by Tukey’s post hoc test, to identify specific group differences. After that, the data were conducted analysis in GraphPad Prism software version 9.0 (GraphPad Software, La Jolla, CA, USA). A p-value of less than 0.05 was regarded as statistically significant.

Results

Characteristics of the polysaccharide extract. The polysaccharide extract of C. lentillifera (sea grape) contained approximately 58.52% total sugar, 11.61% protein, 8.44% sulfate, and 2.62% uronic acid by weight, including a total phenolic content of 7.33 mg GAE/g extract (Table I). Some studies have demonstrated that sulfate in polysaccharides from C. lentillifera is associated with enhanced immunostimulatory activity and reduced lipase activity (23, 24). Moreover, Ismail and Kanaan (25) reported that sulfate and uronic acid in seaweed extracts have a combined effect on antioxidant and antiproliferative activities.

View this table:
Table I.

Chemical composition of the polysaccharide extract of C. lentillifera (sea grape) presented as mean ± SD (n=3).

Analysis of the monosaccharide composition found that the extract consisted of four neutral sugars. The major component was mannose, accounting for 42.49% of the total sugars, followed by galactose at 33.45% and xylose at 17.88%, while L-arabinose accounted for less than 5% of the total sugars (Figure 2 and Table I). This is in line with a study by Le et al. (26), who found mannose was a major component in the C. lentillifera polysaccharide extract. However, the amounts of polysaccharide compositions and types of monosaccharides in the extract might vary depending on the climatic conditions during cultivation and the methods of extraction, separation, and purification.

Figure 2:
Figure 2:

HPLC chromatogram of the monosaccharides in the polysaccharide extract from C. lentillifera. The main monosaccharides identified were xylose, galactose, L-arabinose, and mannose, with retention times of 13.99, 14.75, 15.68, and 16.81 min, respectively. HPLC: High-performance liquid chromatography.

Sea grape treatment induced cytotoxicity and significantly upregulated stemness-related transcription factors in DP cells. Firstly, the cytotoxicity of sea grape in DP cells was assessed using the MTT assay. The results indicated that treatment with sea grape (0-400 μg/ml) did not significantly affect the cell viability compared to untreated control DP cells (Figure 3A). To investigate the impact of sea grape on stemness in DP cells, the cells were treated with 0-50 μg/ml of sea grape for 24 h. The mRNA expression levels of stem cell transcription factors were measured using RT-qPCR, and protein levels were analyzed via immunofluorescence. The mRNA expression levels of the stem cell transcription factors OCT4, NANOG, and SOX2 showed a 20-, 35-, and 15-fold increase, respectively, in DP cells treated with 25 μg/ml of sea grape, as determined by RT-qPCR (Figure 3B).

Figure 3:
Figure 3:
Figure 3:

Cell viability and stemness properties of Dermal Papilla (DP) cells treated with sea grape. (A) DP cells were treated with sea grape (0-400 μg/ml) for 24 h and measured for cell viability. The results indicated that treatment with sea grape (0-400 μg/ml) did not significantly affect the cell viability. (B) mRNA expression levels of the stem cell transcription factors were measured. The mRNA expression levels of the stem cell transcription factors OCT4, NANOG, and SOX2 showed a 20-, 35-, and 20-fold increase, respectively, in DP cells treated with 25 μg/ml of sea grape (C, D) Fluorescence intensity of stem cell transcription factors (OCT4, NANOG, and SOX2) and stem cell markers (CD133, and CD44). The protein expression of stemness transcription factors (OCT4, NANOG, and SOX2) and stemness (CD133 and CD44) in DP cells were significantly increased. (E) Protein expression level of stemness markers. The western blot analysis revealed increased protein expression levels of OCT4, SOX2, CD133, and CD44 proteins. Data are presented as mean±standard deviation (SD) (n=3). Multiple comparisons were made using one-way ANOVA, followed by Tukey’s post hoc test. Significance is indicated as *p<0.05, **p<0.01, and ***p<0.001, compared to untreated control cells.

Immunofluorescence analysis revealed a significant increase in the protein expression of OCT4, NANOG, and SOX2 in DP cells treated with 50 μg/ml of sea grape (Figure 3C). Furthermore, the expression of stem cell markers CD133 and CD44 was found to be 1.5-fold higher in sea grape-treated DP cells, as observed through immunofluorescence (Figure 3D).

DP cells were then exposed to different concentrations of sea grape extract (0-50 μg/ml) for 24 h, followed by western blot analysis to assess the expression levels of OCT4, SOX2, CD133, and CD44 proteins. As shown in Figure 3E, treatment with sea grape significantly enhanced the expression of stem cell markers and transcription factors compared to untreated control cells. These results indicate that sea grape may enhance stemness characteristics.

Sea grape exhibited cytotoxic effects and significantly increased the expression of stemness-related transcription factors in bone marrow-derived UE6E7T-3 cells. Firstly, the cytotoxicity of sea grape in UE6E7T-3 cells was assessed using the MTT assay. The results showed that sea grape treatment did not significantly affect cell viability up to a concentration of 100 μg/ml, compared to untreated cells (Figure 4A). To explore the effect of sea grape on stemness in UE6E7T-3 cells, the cells were treated with sea grape (0-50 μg/ml) for 24 h. The mRNA expression levels of stem cell transcription factors were evaluated using RT-qPCR, and protein expression was analyzed via immunofluorescence.

Figure 4:
Figure 4:
Figure 4:

Cell viability and stemness properties of sea grape treated UE6E7T-3 cells. (A) UE6E7T-3 cells were treated with sea grape extract (0-400 μg/ml) for 24 h, and cell viability was assessed. Results showed that sea grape treatment (0-100 μg/ml) had no significant effect on cell viability. (B) Cells were treated with sea grape extract (0, 25, 50 μg/ml), and mRNA expression of stem cell transcription factors was quantified. Treatment with 50 μg/ml sea grape increased Oct4, Nanog and Sox2 mRNA expression. (C, D) Immunofluorescence analysis showed a significant increase in stemness transcription factors (OCT4, NANOG, SOX2) and markers (CD133, CD44) in UE6E7T-3 cells. (E) The protein expression level of stemness markers assessed by western blot revealed increased protein levels of OCT4, SOX2, CD133, and CD44 proteins in the treated cells. Data are presented as mean±standard deviation (SD) (n=3). Multiple comparisons were made using one-way ANOVA, followed by Tukey’s post hoc test. Significance is indicated as *p<0.05, **p<0.01, and ***p<0.001, compared to untreated control cells.

The mRNA levels of the stem cell transcription factors OCT4, NANOG, and SOX2 increased by 1.5-, 1.5-, and 2.5-fold, respectively, in cells treated with 50 μg/ml of sea grape, as measured by RT-qPCR. However, no significant changes in mRNA levels were observed in cells treated with 25 μg/ml of sea grape (Figure 4B). Immunofluorescence analysis revealed a significant increase in NANOG protein expression, with a 6-fold increase at 25 μg/ml and an 8-fold increase at 50 μg/ml in sea grape-treated cells (Figure 4C). Additionally, the protein levels of OCT4 and SOX2 were significantly increased at the 50 μg/ml dose (Figure 4C).

Furthermore, immunofluorescence analysis showed a significant increase in the expression of stem cell markers CD133 and CD44 in sea grape-treated UE6E7T-3 cells (Figure 4D). Additionally, UE6E7T-3 cells were treated with varying concentrations of sea grape extract (0-50 μg/ml) for 24 h, and the expression levels of stem cell transcription factors (OCT4, SOX2) and stemness markers (CD133, CD44) were evaluated using western blot analysis. The findings showed that treatment with 50 μg/ml of sea grape extract led to a 1.5-fold increase in the expression of CD44 and SOX2 compared to untreated control cells (Figure 4E). These results suggest that sea grape may enhance stemness features in UE6E7T-3 cells, highlighting its potential role in skin rejuvenation.

The sea grape treatment affected the viability of human melanoma cells and decreased melanin content in human melanogenesis-related cells. A cell viability assay was performed to assess the cytotoxicity of sea grape in human G361 melanoma cells. G361 cells were treated with varying concentrations of sea grape (0-100 μg/ml) for 24, 48, and 72 h. No significant change in cell viability was observed until the lowest concentration of 20 μg/ml (Figure 5A). Figure 5C shows a significant decrease in cellular melanin content after incubation with sea grape concentrations ranging from 0 to 20 μg/ml for 72 h. The results demonstrated a significant anti-melanogenic effect at all time points measured for melanin content.

Figure 5:
Figure 5:

Sea grape affects cytotoxicity and inhibits melanin production in human G361 melanoma cells. (A) The melanoma cells were treated with (0-100 μg/ml) sea grape for 24, 48, and 72 h. No significant change in cell viability was observed until 20 μg/ml. (B) Treatment of the cells with sea grape at 0, 10, and 20 μg/ml resulted in the inhibition of melanogenesis. (C) The G361 cells were treated with sea grape for 24, 48, and 72 h and the cellular melanin content was determined. The results show a significant decrease in the melanin content. (D) Cellular tyrosinase activity was measured in sea grape extract-treated cells, showing a significant reduction at all time points. Data are presented as mean±standard deviation (SD) (n=3). Multiple comparisons were made using one-way ANOVA, followed by Tukey’s post hoc test. Significance is indicated as *p<0.05 and ***p<0.001, compared to untreated control cells.

Tyrosinase enzymes play a crucial role in regulating melanogenesis in melanoma cells (27). To assess the effect of sea grape on tyrosinase activity, a cellular tyrosinase assay was conducted. The impact of sea grape on tyrosinase activity was examined using a cell-based model, where the enzyme tyrosinase catalyzed the conversion of the substrate L-Dopa to dopachrome. As shown in Figure 5D, protein lysates from sea grape-treated human G361 melanoma cells exhibited reduced tyrosinase activity at 24, 48, and 72 h. A significant reduction in cellular tyrosinase activity was observed at 24 h in sea grape-treated human G361 cells.

Discussion

Skin aging refers to the progressive decline in skin quality over time, driven by the combined effects of chronological aging, sun exposure, hormonal changes, and environmental influences. While various treatments exist for skin rejuvenation, natural product compounds play a significant role in addressing skin aging (28, 29). As a result, numerous researchers have been exploring scientific approaches to delay skin aging and enhance skin rejuvenation. Skin discoloration is a key aspect of aging, often resulting from increased melanin production, which leads to the appearance of dark spots. The formation of melanin is influenced by tyrosinase activity during melanogenesis, playing a significant role in the aging process (30).

Skin disorders involving abnormal pigmentation accumulation are challenging to address due to unclear etiologies and unknown pathological causes. The connection between skin aging and pigmentation abnormalities is often linked to inherited conditions associated with premature aging (31). As a result, many researchers focus on investigating ways to reduce melanogenesis in aging-related conditions, particularly through the protective use of natural products, especially those of marine origin. Fucoidan extracted from marine brown seaweed and Turbinaria conoides (brown alga) inhibits tyrosinase activity and demonstrates significant skin whitening activity (32, 33).

Previous reports indicate that various secretomes from different types of stem cells, such as conditioned medium (CM) from human adipose-derived stem cells (ADSCs), human umbilical cord blood (hUCB), human placental stem cells (hPSCs), and human neural stem cells (NSCs), regulate melanin production through various pathways (9, 10). The CM primarily involves cytokines, chemokines, growth factors, metabolites, bioactive lipids, and vesicles or exosomes that may function in an autocrine or paracrine manner by inhibiting melanin synthesis in the skin (9, 10).

Our research demonstrated that treatment with the polysaccharide form of sea grape extracts led to a 20-, 35-, and 15-fold increase in the mRNA expression levels of stem cell transcription factors OCT4, NANOG, and SOX2, respectively, in DP cells (Figure 3B). Additionally, in sea grape-treated bone marrow-derived UE6E7T-3 cells, the mRNA expression of OCT4, NANOG, and SOX2 increased by 1.5-, 1.5-, and 2.5-fold, respectively (Figure 4B). The protein expression levels of stem cell markers (CD133, CD44) and transcription factors (OCT4, NANOG, SOX2) were significantly elevated in sea grape-treated DP and UE6E7T-3 cells.

Furthermore, sea grape treatment significantly reduced melanin synthesis in human melanoma cells (G361) after 72 h (Figure 5B, C). These findings suggest that the phenolic compounds in the sea grape extract (Table I) may contribute to the observed reduction in melanin synthesis. A previous study by Chaiklahan et al. (34) identified quercetin and quercetagetin as predominant phenolic constituents in sea grape extracts. Both compounds have been reported to inhibit tyrosinase activity and suppress melanogenesis (35-37), supporting their potential role in the observed reduction of melanin synthesis.

L-DOPA and L-tyrosine, beyond being melanogenesis substrates, also act as bioregulatory agents that both initiate and regulate melanogenesis and other cellular functions, with the pathway capable of self-regulation and modulating melanocyte activity through its proteins and intermediates (27, 38, 39). Based on our findings, sea grape treatment markedly decreased tyrosinase activity in human melanoma cells at 24, 48, and 72 h (Figure 5D).

Conclusion

In conclusion, C. lentillifera, or sea grape, has demonstrated the ability to decrease melanin synthesis and enhance regenerative markers in stem cells. This presents potential for creating innovative products aimed at melanin regulation in melanoma cells and skin rejuvenation, due to its ability to boost stem cell markers in human DP cells and UE6E7T-3 cells derived from the bone marrow.

Acknowledgements

Not applicable.

Footnotes

  • Authors’ Contributions

    Conceptualization: PC. Methodology: PC, RC. Validation: PC, RC. Formal analysis: ZZE, PC. Investigation: ZZE, PC. Resources: NC, TS, RC. Writing-original draft preparation: RC, ZZE, BI. Writing–review and editing: PC. Supervision: PC. All Authors have read and agreed to the final version of the manuscript.

  • Conflicts of Interest

    All Authors declare that there are no conflicts of interest.

  • Funding

    This project was funded by National Research Council of Thailand (NRCT): NRCT Research Chair Grant (Grant number: N42A670596).

  • Artificial Intelligence (AI) Disclosure

    During the preparation of this manuscript, a large language model (ChatGPT, OpenAI) was used solely for language editing and stylistic improvements in select paragraphs. No sections involving the generation, analysis, or interpretation of research data were produced by generative AI. All scientific content was created and verified by the authors. Furthermore, no figures or visual data were generated or modified using generative AI or machine learning-based image enhancement tools.

  • Received May 29, 2025.
  • Revision received August 11, 2025.
  • Accepted September 10, 2025.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY-NC-ND) 4.0 international license (https://creativecommons.org/licenses/by-nc-nd/4.0).

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Polysaccharides Derived from Caulerpa lentillifera (Sea Grape) Inhibit Melanogenesis in Human Melanoma Cells and Promote Stemness in Stem Cells
RATANA CHAIKLAHAN, ZIN ZIN EI, NATTAYAPORN CHIRASUWAN, THANYALAK SRINORASING, BHURICHAYA INNETS, PITHI CHANVORACHOTE
In Vivo Nov 2025, 39 (6) 3258-3270; DOI: 10.21873/invivo.14125
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