Fetal bovine serum (FBS) is widely used for cell culture media, especially its function as a growth supplement with high growth-promoting factors. An optimal culture medium is needed to increase protein transduction. Unfortunately, FBS reported as media contained protease and contaminated with pathogen microorganisms from an animal host. Fibroblast preputium cell is easy to culture and can be a good model for assessing the medium culture system. In this study, FBS was substituted with honey and royal jelly to find an alternative FBS. This study aimed to determine the effectivity of serum-free DMEM medium with honey from Tetragonula sp. and royal jelly from Apis mellifera (Ceiba pentandra) on the proliferation of fibroblasts preputium cells. The research design used true experimental methods. Samples were taken from healthy people. Fibroblast cells were cultured with various concentrations of honey and royal jelly (0.1%, 1%, 5%). The best result of those various concentrations continued until 9 days with continuous checking in every three days measured with Microtetrazolium (MTT) assay test. Fibroblast cells cultured in Tetragonula sp. honey and royal jelly Apis mellifera (Ceiba pentandra) 0.1% medium had a significant difference, with proliferation higher than 1% (p = 0,000) and 5% (p = 0,000), but did not exceed proliferation with FBS addition medium. Next, cells in DMEM medium with Tetragonula sp. honey and Apis mellifera royal jelly (Ceiba pentandra) 0.1% on the 3rd, 6th, and 9th (p = 0,000; p = 0,000; p = 0,000) had not similar growth to the standard medium with FBS. However, the growth on the 9th day had a significant difference with the DMEM medium without FBS. High sugar in honey can inhibit fibroblast cell proliferation. The addition of other components as needed to optimize proliferation in honey and royal jelly medium. Isolation of active ingredients in honey and royal jelly can function as an alternative development of an effective and safe substitute for FBS.

ABSTRAK Nutrient agar merupakan jenis media umum yang sering digunakan pada laboratorium, harga media ini relatif mahal. Nutrient agar berbentuk padat, terdiri atas campuran ekstrak daging, pepton serta agar. Ikan teri jengki dan ikan layang deles jumlahnya melimpah di Indonesia. Ikan teri jengki (Stolephorus indicus) mengandung protein 51.54%, sementara ikan layang deles (Decapterus Macrosoma) mengandung protein sebesar 57,89%. Escherichia coli digunakan sebab bakteri ini merupakan salah satu jenis mikroorganisme kontrol positif yang direkomendasikan berdasarkan Data Sheet Nutrient Agar OXOID. Penelitian ini bersifat eksperimental dengan tiga perlakuan pada masing-masing bahan yaitu variasi massa 3 gram, 4 gram dan 5 gram yang dilakukan di Laboratorium Bakteriologi Jurusan Teknologi Laboratorium Medis Poltekkes Kemenkes Surabaya. Hasil penelitian menunjukkan jumlah koloni rata-rata Escherichia coli yang tumbuh pada kontrol positif sebanyak 118 x 1013, sementara pada media ikan teri jengki variasi massa 3 gram adalah 72 x 1013 CFU/mL, 4 gram sebanyak 85 x 1013 CFU/mL, 5 gram sebanyak 94 x 1013 CFU/mL. Pada media ikan layang deles variasi 3 gram sebanyak 88 x 1013 CFU/mL, 4 gram sebanyak 96 x 1013 CFU/mL, 5 gram 108 x 1013 CFU/mL. Variasi massa yang paling baik dan mendekati hasil kontrol positif (nutriet agar) pada ikan teri jengki dan ikan layang deles adalah variasi massa 5 gram.Kata kunci: ikan teri jengki, ikan layang deles, Escherichia coli, nutrient agar