EVALUATION OF RAT LEYDIG CELL CULTURE COLLECTED WITH NYCODENZ GRADIENT IN PRODUCING TESTOSTERONE IN VITRO

Ekayanti Mulyawati Kaiin, Wahono Esthi Prasetyaningtyas

Abstract


 

The aim of this study was to evaluate the ability of rat’s Leydig cells collected with Nycodenz gradient in producing testosterone in vitro. Leydig cells were collected using 5 column of Nycodenz gradient (4, 8, 10, 12, and 15%) and cells were evaluated regarding its concentration, viability, and purity of Leydig cells. Media used to cultured Leydig cells were Dulbecco’s Modified Eagle’s Medium (DMEM)+10% newborn calf serum (NBCS); DMEM+10% NBCS+2,5 IU/mL human chorionic gonadotrophin (hCG); DMEM+10% NBCS+ 5 µg/mL insulin, 10 µg/mL transferrin, and 5 µg/mL Se (ITS); DMEM+10% NBCS+hCG+ITS at 5% CO2 incubator with temperature of 37.5° C for 3 days. Culture medium was collected every day for testosterone analysis with enzyme-linked immunosorbent assay (ELISA). By adding ITS to the medium, Leydig cells concentration was significantly increased (8.92x106  cells/mL) compared to medium with serum (7.74x106 cells/mL) or hCG (7.68x106 cells/mL) (P<0.05). ITS and hCG in medium significantly increased Leydig cells concentration (10.40x106 cells/mL) at day 3 of culture (P<0.05). The result of parallelism test showed that the assay obtained good validity to measure testosterone concentration in culture medium. Testosterone in medium was detected at 1.80-2.60 ng/mL at day 1 of culture. In conclusion, Leydig cells collected with Nycodenz gradient had no effect to testosterone secretion from Leydig cells in vitro.


Keywords


ELISA; hCG; Leydig cells; ITS; In vitro culture; Leydig cells; Nycodenz; Testosterone

Full Text:

PDF


DOI: https://doi.org/10.21157/j.ked.hewan.v11i4.3984

Refbacks

  • There are currently no refbacks.


Copyright (c) 2018 by author and J. Ked. Hewan

Creative Commons License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

Editorial Office

Jurnal Kedokteran Hewan (Indonesian Journal of Veterinary Sciences)

Faculty of Veterinary Medicine Syiah Kuala University
Jl. Tgk. Hasan Krueng Kalee No. 4, Kampus FKH Unsyiah Darussalam
Banda Aceh 23111, Indonesia
Phone/Fax. +62-651-7551536, CP: +6285260153935; +6281362693330
e-mail: jkh@unsyiah.ac.id; jurnal_khusk@yahoo.com

  

p-ISSN: 1978-225X e-ISSN: 2502-5600


Creative Commons License
J Ked Hewan
is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.