Skripsi

Preliminary Study of Prepuce Skin Cell Culture: Identification of Oct-4 Positive Cells and Ki-67 Positive Cells from Dermis and Hypodermis Layer = Studi Preliminari Kultur Sel dari Preputium: Identifikasi Sel Positif Oct-4 dan Ki-67 di Dermis dan Hipodermis.

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’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 pelletswere 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 7 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 cell were found in dermis sample and there were no ki-67 positive cells. To conclude, more cells are needed to further analyse oct-4 and ki-67 positive cells in dermis and hypodermis of prepuce. Therefore, optimization of isolation and culture method for prepuce is still needed.
Key words : Stem cell, prepuce, culture, isolation, oct-4, ki-67, harvest, immunocytochemistry


Studi tentang sel punca pluripoten di preputium adalah salah satu topik yang belum banyak dilakukan namun sedang berkembang. Beberapa penanda seperti penanda pluripotensi oct-4 dan penanda proliferasi ki-67 telah dilaporkan keberadaannya di preputium. Untuk menumbuhkan sel punca tersebut, teknik isolasi dan kultur yang baik perlu dilakukan. Pengertian lebih dalam mengenai sel punca di preputium dapa tmembuka kemungkinan baru dalam terapi rekonstruksi kulit seperti cangkok kulit. Tujuan riset ini adalah untuk melakukan penetapan metode awal isolasi dan kultur sel dermis dan hipodermis preputium dan mengidentifikasi sel positif oct-4 dan ki-67. Dermis dan hipodermis diisolasi dan dikultur di 12-well plate dan dipindahkan ke 24-well plate. Kultur diobservasi untuk identifikasi sel-sel fibroblastik. Panen sel dilakukan dengan tripsinasi dan sentrifugasi. Pellet difiksasi dengan methanol dan di-mount ke preparat histologi. Preparat diproses immunositokimia dengan antibodi oct-4 dan ki-67 dan diobservasi dibawah mikroskop cahaya. Pada kesimpulannya, dibutuhkan lebih banyak sel untuk analisis sel positif oct-4 dan ki-67. Oleh karena itu, optimasi metode isolasi dan kultur dermis dan hipodermis preputium masih diperlukan.
Kata kunci : Selpunca, preputium, kultur, isolasi, oct-4, ki-67, panen sel, immunositokimia