Geraniin supplementation increases human keratinocyte proliferation in serum-free culture

Main Article Content

Indra Kusuma
Restu Syamsul Hadi


BACKGROUND Various products used in cellular therapy utilize tissue culture techniques requiring keratinocyte culture. An efficient and clinically acceptable keratinocyte culture system requires supplements with mitogenic activity. Geraniin is a phytochemical with the potential as a supplement for expansion culture of keratinocytes. The objective of the present study was to verify the mitogenic activity of geraniin on human keratinocytes. METHODS This was an experimental study using two samples of human foreskin obtained by circumcision of a male child. Epidermal keratinocytes were isolated from the foreskin samples and were divided into paired groups, comprising intervention and control groups. The intervention groups were cultured with geraniin supplementation, whereas the control groups with standard supplements, without the addition of geraniin. Mitochondrial activity of the cells was evaluated by means of the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium-bromide (MTT) proliferation assay. Absorbance values in each of the groups was measured at 450 nm. Data analysis was performed with the paired t-test. RESULTS Geraniin supplementation significantly increased the keratinocyte proliferation rates at dosages of 0.8 to 3.1 μM. An increase of 57% in the proliferation rate was obtained at a dosage of 1.6 μM, while at a dosage of 12.5 μM toxic effects were starting to appear. Geraniin presumably causes increased cellular energy status, resulting in increased proliferation rates. CONCLUSION The findings in this study provide evidence in support of the utilization of geraniin as a supplement for expansion culture of keratinocytes. Further studies may presumably identify the molecules acting as geraniin receptors and the intracellular mechanisms underlying the increase in proliferation rates.

Article Details

How to Cite
Kusuma, I., & Hadi, R. S. (2013). Geraniin supplementation increases human keratinocyte proliferation in serum-free culture. Universa Medicina, 32(1), 3–10. - 10
Review Article


Corte PD, Verween G, Verbeken G, Rose T, Jennes S, Coninck AD, et al. Feeder layer- and animal product-free culture of neonatal foreskin keratinocytes: improved performance, usability, quality and safety. Cell Tissue Bank 2012;13:175-

Coolen NA, Verkerk M, Reijnen L, Vlig M, Bogaerdt AJ, Breetveld M, et al. Culture of keratinocytes for transplantation without the need of feeder layer cells. Cell Transplant 2007;16:649-61.

Grinnell KL, Bickenbach JR. Skin keratinocytes pre-treated with embryonic stem cell-conditioned medium or BMP4 can be directed to an alternative cell lineage. Cell Prolif 2007;40:685-705.

Grinnell KL, Yang B, Eckert RL, Birkenbach JR. Diferentiation of mouse interfollicular keratinocytes by the embryonic transcription factor Oct-4. J Investig Dermatol 2006;127:372-

Sun X, Fu X, Han W, Zhao Y, Liu H, Sheng Z. Dediferentiation of human terminally dfferentiating keratinocytes into their precursor cells induced by basic fibroblast growth factor.

Bio Pharm Bull 2011;34:1037-45.

Aasen T, Belmonte J. Isolation and cultivation of human keratinocytes from skin or plucked hair for the generation of induced pluripotent stem cells. Nat Prot 2010;5:371-82.

Ohmine S, Squillance K, Hartjes K, Deeds M, Amstrong A, Thatava T, et al. Reprogrammed keratinocytes from elderly type 2 diabetes patients suppress senescence to acquire induced pluripotency. Aging 2012;4:60-73.

Kalyanaraman B, Boyce ST. Wound healing in athymic mice with engineered skin subtitutes fabricated with harvested from automated bioreactor. J Surg Res 2009;152:296-302.

Setyowati FM. Etnofarmakologi dan pemakaian tanaman obat suku dayak tunjung di Kalimantan Timur. Media Litbang Kesehatan 2010;20:104-12.

Yuwono S. Pengaruh pemberian meniran pada hati mencit yang diberi CCl4. CDK 2010;177:278-

Dalimartha S. Atlas tumbuhan obat Indonesia. Jakarta: Trubus Agriwidaya; 2003.

Agyare C, Lectenberg M, Deters A, Petereit F, Hensel A. Ellagitannins from Phyllantus muellerianus (Kuntze) Excell: Geraniin and furosin stimulate cellular activity, differentiation, and

collagen synthesis of human skin keratinocytes and dermal fibroblasts. Phytomedicine 2011;18: