Whitening Effect of Eucheuma cottonii Extract

in Phorbol-12-myristate-13-acetate induced
Melanocytes

Monica Drestia
2011-080-074
(Costin & Hearing 2007).
→ The Factors...

(Costin & Hearing 2007).

 (Selvan et al. 2014).

B16F10 Murine
Melanocytes
 Methods
Sample Extraction

Cell Cultivation

Melanogenesis Induction & Sample

Treatment
Measurement of Tyrosinase Activity

Cytotoxicity Assay

RNA Isolation & qRT-PCR

Sample Extraction

+ → →

←
(Hindritiani et al. 2013).

Cell Cultivation

→
Melanogenesis Induction

Sample Treatment

(Lee et al. 2010).

Cytotoxic Assay
37oC
3 hr

+ → 590 nm

MTT reagent
Treated cells (20 µL)
(Hindritiani et al. 2013).

Measurement of Tyrosinase Activity

37oC
1 hr

→ →
+ L-DOPA 475 nm

Treated cells,
lysed
RNA Extraction & qRT-PCR

→
+ Trizol

treated cells
Kappa SYBR FAST One-Step qRT-PCR

 Primer :
- GAPDH
- Mus musculus microphthalmia-associated
transcription factor (MITF)
 Results
100
90
Cell Viability (%)

80
70
60 E. cottonii
50
Ascorbic
40
Acid
30
20
10
0
0 5 10 25 50 100 150 250

Sample concentration (µg mL-1)

Figure 1 Cell viability percentage of B16F10 cells after PMA

induction and 24h incubation of samples treatment.
 Results
100
90
Tyrosinase activity

80
(% control)

70 E. cottonii
60 Linear (E. cottonii)
50
40 Ascorbic Acid
30
20
10
0
0 5 10 25 50 100 150 250

Sample concentration (µg mL-1)

Figure 2 The effect of E. cottonii extract on the tyrosinase activity

of treated B16F10 cells.
 Results
100
90
Relative MITF gene

80
expression (%)

70
60 E. cottonii
50
40
Ascorbic acid
30
20
10
0
0 5 10 25 50 100 250

Sample concentration (ug mL-1)

Figure 3 The effect of E. cottonii extract on the mRNA expression of

melanogenic transcription factor MITF of treated B16F10 cells.
 Conclusion

• E. cottonii extract significantly reduced

tyrosinase activity and MITF expression level
of the cells without any significant cytotoxic
activity on cell proliferation
• E. cottonii extract can be considered as skin
whitening agent
 References
Costin GE, Hearing VJ. 2007. Human skin pigmentation: melanocytes modulate
skin color in response to stress. FASEB.21:976-994. 
Hindritiani R, Dhianawaty D, Sujatno M, Sutedja E, Setiawan. 2013. Penurunan
aktivitas tirosinase dan jumlah melanin oleh fraksi etil asetat buah malaka
(Phyllantus emblica) pada mouse melanoma B16 cell-line. MKB.45(2):118-124.
Lee CW, Kim HS, Kim JW, Yoon JH, Cho Y, Hwang JK. 2010. Inhibitory effect
of panduratin a isolated from Kaempferia panduarata Roxb. on melanin
biosynthesis. Phytother Res. 24:1600–1604.
Selvan BK, Piriya PS, Chandrasekhar M, Vennison SJ. 2014. Macro algae
(Eucheuma cottonii and Sargassum sp) are reservoirs of biodiesel and bioactive
compounds. J Chem Pharm. 2:62-70.
Song YS, Balcos MC, Yun HY, Baek KJ, Kwon NS, Kim MK, Kim DS. 2015.
ERK activation by fucoidan leads to inhibition of melanogenesis in Mel-Ab
cells. Korean J Physiol Pharmacol. 19:29-34.
 Many thanks to:
• Lord Jesus Christ and Mother Mary
• Yonsei University
• Faculty of Biotechnology Atma Jaya Catholic University 0f Indonesia
• Ms.Yanti, PhD as my advisor
• Mr. Adi Yulandi S.Si, M.T as today’s moderator
• Mom, Dad, and my family
• The Lovely Skin Team (Patrice, Keren, Margareth, Halim, Alwi)
• PSSP Team (Bu Diah, Bu Silmi)
• Yudith and Willchan as today’s awesome partner
• Kak Rika,Teteph, Kimberly, Oline Anggasta, Edu
• Penghuni lab kuljar (Grace, Disa, Kent, Boni, Dira, Laudus, Yunita, Edbert,
Tjong, Sur, Natan), Odi, Merci
• Penghuni lab mikro (*Vege, Mike, Pau), penghuni lab biokim (*Kak Fani,
Stella, Olga, Ocil, Hux, Debi, geng bioplastic)
• FTb 2011 family, FTb 2012, FTb 2013, FTb 2014
• Technisians
• All of today’s seminar attendants 