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Abstrak :
ABSTRAK
Studi tentang sel punca pluripoten di preputium adalah salah satu topik yangbelum banyak dilakukan namun sedang berkembang. Beberapa penanda sepertipenanda pluripotensi oct-4 dan penanda proliferasi ki-67 telah dilaporkankeberadaannya di preputium. Untuk menumbuhkan sel punca tersebut, teknikisolasi dan kultur yang baik perlu dilakukan. Pengertian lebih dalam mengenai selpunca di preputium dapa tmembuka kemungkinan baru dalam terapi rekonstruksikulit seperti cangkok kulit. Tujuan riset ini adalah untuk melakukan penetapanmetode awal isolasi dan kultur sel dermis dan hipodermis preputium danmengidentifikasi sel positif oct-4 dan ki-67.Dermis dan hipodermis diisolasi dan dikultur di 12-well plate dan dipindahkan ke24-well plate. Kultur diobservasi untuk identifikasi sel-sel fibroblastik. Panen seldilakukan dengan tripsinasi dan sentrifugasi. Pellet difiksasi dengan methanol dandi-mount ke preparat histologi. Preparat diproses immunositokimia denganantibodi oct-4 dan ki-67 dan diobservasi dibawah mikroskop cahaya.Pada kesimpulannya, dibutuhkan lebih banyak sel untuk analisis sel positif oct-4dan ki-67. Oleh karena itu, optimasi metode isolasi dan kultur dermis danhipodermis preputium masih diperlukan.

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ABSTRACT
The study on pluripotent stem cell in prepuce is one of the topic on stem cell that is not yet common but rapidly developing. Several markers such as Oct 4 for pluripotency and ki 67 for proliferation have been reported in prepuce. To generate these stem cells, careful isolation and cell culture are performed. Better understanding of stem cells in prepuce can open more possibilities in skin regeneration and skin reconstruction treatment such as skin graft. This research aims to perform initial establishment method for isolation and culture of prepuce skin rsquo s dermis and hypodermis and to identify oct 4 and ki 67 positive cells from the culture derived cells.The dermis and hypodermis of prepuce were isolated and cultured in 12 well plate for 7 days and transferred into 24 well plate. The culture was observed for fibroblastic like cells. The cells were harvested using trypsinization and centrifugation. The pellets were fixated on methanol to be mounted on histological slides. Immunocytochemistry using oct 4 and ki 67 antibody were performed on the samples. The samples were observed under light microscope.Fibroblastic like cells were found in one of the culture on the 7th day. Dermis culture has more pellet of harvested cells compared to hypodermis culture. Histological slides examination revealed few number of cells and debris can be found. Oct 4 positive cells were found in dermis sample and there were no ki 67 positive cells.

Abstrak :
Latar Belakang: Kondiloma akuminata adalah infeksi menular seksual yang disebabkan oleh Human Papilloma Virus HPV . Epitel mukosa pada kulit prepusium penis yang utuh pada lelaki tidak disunat diketahui sebagai lokasi predileksi lesi proliferatif Kondiloma akuminata. Fenomena ini dipengaruhi oleh HPV yang menyukai permukaan yang lembab. Diperlukan studi lebih lanjut mengenai kapasitas proliferasi keratinosit basal pada epitelium prepusium penis yang diduga merupakan faktor lain yang berpengaruh pada lokasi predileksi Kondiloma akuminata. Metode: Sampel kulit prepusium didapat dari 10 partisipan acara sunatan massal. Sampel melalui proses histoteknik dan diwarnai dengan pewarnaan Hematoxylin-Eosin dan immunohistokimia. Preparat diobservasi menggunakan mikroskop Olympus untuk dilakukan penghitungan manual pada keratinosit basal yang positif Ki-67. Hasil dianalisis menggunakan Microsoft Excel 2010. Hasil: Dari 10 sampel, terdapat 2 sampel dengan histologi kulit prepusium penis yang lengkap, 5 sampel yang hanya memiliki epitel mukosa dalam dan 3 sampel yang hanya memiliki epithel kutan luar. Rata-rata dari basal keratinosit yang positif Ki-67 pada epitel mukosa adalah 5.9 sel, sedangkan epitel kutan luar memiliki rata-rata 3.6 sel. Kesimpulan: Epitel mukosa bagian dalam memiliki lebih banyak keratinosit basal yang berproliferasi dibandingkan dengan sel di epitel kutan bagian luar dari kulit prepusium penis. Jumlah sel yang lebih banyak berproliferasi diduga merupakan faktor yang berpengaruh pada lokasi predileksi Kondiloma akuminata. ......Background: Condyloma acuminata is a sexually transmitted infection caused by Human Papilloma Virus HPV . In uncircumcised men with intact preputial skin of penis, mucosal surface is the predilection site of proliferative lesion in Condyloma acuminata. This is explained by HPV preference towards moist surface. Further investigation is necessary to understand the capacity of basal keratinocyte of penile preputial skin epithelium to proliferate, as it is suspected as a factor contributing to the predilection site of Condyloma acuminata. Method: Preputial skin samples were obtained from 10 participants in mass circumcision event. Samples underwent histotechnique process and stained by Hematoxylin Eosin and immunohistochemistry. Microscopic slides were observed under Olympus microscope to allow manual counting of Ki 67 positive cell. The data collected was analyzed using Microsoft Excel 2010. Results: From 10 samples, there were 2 samples with complete penile preputial skin histology, 5 samples with inner mucosal epithelium only and 3 samples with outer cutaneous epithelium only. Mean of Ki 67 positive basal keratinocyte in inner mucosal epithelium was 5.9 cells, while outer cutaneous epithelium was 3.6 cells. Conclusion: Inner mucosal epithelium showed more Ki 67 positive cells compared to outer cutaneous epithelium. More actively proliferating cell in the mucosal epithelium may serves as factor influencing the predilection site of Condyloma acuminata.