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Journal : HAYATI Journal of Biosciences

3D Culture Cells Technique for Exosomes Isolation of HEK293 and its Application on WiDr Cells Audina, Mia; Mariya, Silmi; Zaelani, Bella Fatima Dora; Yuliana; Darusman, Huda Shalahudin
HAYATI Journal of Biosciences Vol. 31 No. 6 (2024): November 2024
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.31.6.1173-1180

Abstract

Three-dimensional (3D) culture is a technique commonly utilized in bioprocessing and biomedical research. Exosomes have been investigated as carriers for medications in numerous studies employing 3D culture methodologies. The objective of this research is to employ 3D cell culture for the isolation and treatment of exosomes targeting colon cancer cells. The isolation of exosomes obtained from HEK293 cells was conducted through the ultracentrifugation technique. Subsequently, exosome treatment was administered to WiDr cells at concentrations of 3.5 µg/ml, 7 µg/ml, and 14 µg/ml.The validation of molecular markers of exosomes (CD9 and CD81), along with BAX, BCL-2, and CD133, was performed using qRT-PCR. The findings revealed the successful isolation of exosomes derived from HEK293 cells, which exhibited the expression of markers CD9 and CD81. Furthermore, the expression of BAX and BCL-2 indicated the potential of exosomes to induce apoptosis, while the expression level of CD133 decreased with treatment at varying concentrations. These results suggest that exosome treatment has the capability to impede the proliferation of WiDr cells and reduce the expression of CD133, thereby signifying the potential application of exosomes as an in-vitro model for investigating cancer therapy in the future.
Cytotoxicity of Ethyl Acetate Extract of Avocado Seeds (Persea americana Mill.) on Mouse Melanoma B16F10 Cell Line Sebayang, Shalina; Mariya, Silmi; Rayendra, Raendi; Wientarsih, Ietje; Priosoeryanto, Bambang Pontjo
HAYATI Journal of Biosciences Vol. 32 No. 2 (2025): March 2025
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.32.2.405-413

Abstract

Artificial whitening agents available in the market can cause damage to melanocytes; therefore, a safe and naturally derived skin-whitening agent is needed. Avocado seeds (Persea americana Mill.) are often discarded as waste but possess numerous health benefits. One of the compounds in avocado seeds is catechin, a flavonoid metabolite. This compound has been reported to exhibit antioxidant activity and inhibit tyrosinase to prevent melanin formation, making it suitable for skin-whitening applications. This study aimed to conduct phytochemical screening, assess antioxidant activity using the DPPH method, analyze catechin content using HPLC, and perform cell viability tests using the MTT method from the ethyl acetate extract of avocado seeds. Phytochemical screening has revealed the presence of flavonoids, saponins, and steroids. The DPPH assay yielded an IC50 value of 89.47±0.73 ppm, indicating an intense antioxidant activity, and HPLC detected 0.09% catechin. The MTT test results yielded cell viability percentages ranging from 80-100% at test concentrations of 0.75-25 ppm, with an IC50 value of 61.7 ppm. In conclusion, our results indicate that the ethyl acetate extract from avocado seeds affects cell viability without toxicity, warranting further testing for tyrosinase inhibition in the mouse melanoma B16F10 cell line.
Evaluating the Cytotoxic Effects of Ethanol and n-Hexane Extracts from Black Cumin Seeds (Nigella sativa) on B16F10 Mouse Melanoma Cells : A Preliminary Investigation into Vitiligo Treatment Pratiwi, Dian; Mariya, Silmi; Rayendra, Raendi; Setiyono, Agus
HAYATI Journal of Biosciences Vol. 32 No. 1 (2025): January 2025
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.32.1.203-211

Abstract

Black cumin (Nigella sativa) is a herbal plant that has been cultivated locally in Indonesia and is traditionally used for various diseases. Thymoquinone, one of the main components, is rich in biological activity. In several countries, topical application of its oil on human skin with vitiligo can stimulate skin repigmentation with minimal side effects. This study aims to determine the viability of B16F10 melanoma mouse cells against ethanol and n-hexane extracts of black cumin seeds through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The ethanol extract (EE) yield was 14.306%, and the n-hexane extract (NHE) was 7.442%. Phytochemical screening of EE detected flavonoids, alkaloids, saponins, and steroids, and High-performance liquid chromatography (HPLC) detected 0.040% thymoquinone. The MTT test showed cell viability was >100% from EE at all treatment concentrations, namely 0.75-100 ppm and only 0.75-6.25 ppm for NHE. In conclusion, this study indicates that 96% EE of Nigella sativa is less toxic than NHE on B16F10 mouse melanoma cells and has potential as an alternative treatment for vitiligo, which needs to be proven in further research.