ABSTRAK
Sampah medis berupa prepusium sangatlah mudah di peroleh di Indonesia. Sel-selyang di dapat dari enam sampel di duga memiliki kapasitas pluripotensi dan dapatberdiferensiasi ke lini lain untuk pengobatan secara medis. Pada experiment ini, selkeratinosit di peroleh dari epidermis prepusium, sel tersebut di ambil mengunakanlarutan enzymatik dispase dan tripsin masing-masing di inkubasi semalaman dengansampel. Kemudian sel-sel tersebut di kultur dengan DMEM tinggi glukosa untukmeningkatkan jumlah sel. Setelah di kultur, sel-sel tersebut di ambil untukpengecekan immunositokimia (ISK) Oct-4, hal ini di karenakan Oct-4 adalahpenanda kapasitas pluripotensi. Sample lainnya tetap di kultur untuk eksperimenkonfluensi/differensiasi spontan selama 14 hari. Riset ini telah sukses menggunakanmedium kultur alternatif dan berbiaya efektif untuk menkultur keratinosit. Bahanmedium tersebut yakni; DMEM tinggi glukosa, PRP 10%, heparin 1%, FBS 10%,penstrep 1%, dan fungizone 1%. Hasil dari pada ISK tersebut adalah positif parsialdengan nukleus keratinosit yang terwarnai coklat tua pada lima lapang pandangberkekuatan tinggi dari setiap sampel. Namun, analisis diferensiais spontanmenggunakan alcian blue menunjukkan hasil negative dengan tidak adanyaperubahan dari lini keratinosit ke kondrosit
ABSTRACT
The medical waste of preputial skin is easily obtained in Indonesia. The cells isolatedfrom six samples are expected to have pluripotency and able to differentiate to otherlineage for medical treatment. This research uses the epidermal layer of preputial skinto obtain keratinocytes, this cells are taken using dispase and trypsin solution overnightrespectively. Then, the keratinocytes are subsequently cultured using DMEM completehigh glucose to increase the number of cells. The cultured cells are then taken forimmunocytochemistry (ICC) of Oct-4, since it is the marker of pluripotency. The otherhalf of cultured samples are continued for over confluency analysis for fourteen daysto observe spontaneous differentiation. This research has successfully used analternative and cost effective culture medium for keratinocytes. It consist of DMEMhigh glucose, PRP 10%, heparin 1%, FBS 10%, penstrep 1%, and fungizone 1%. Theresult of ICC is partially positive with keratinocytes nuclear being stained dark brownin five hpf from each sample. However, spontaneous differentiation analysis usingalcian blue shows negative result of chondrogenic formation from keratinocytes |
S-Jaspal Singh.pdf :: Unduh