TITLE:
Development of in vitro culture of rat Leydig cells after purification with Nycodenz gradient
AUTHORS:
Ekayanti Mulyawati Kaiin, Ita Djuwita, Tuty Laswardi Yusuf, Mohamad Agus Setiadi
KEYWORDS:
Leydig; In Vitro; Rat; Nycodenz; hCG; ITS; Testosterone
JOURNAL NAME:
Open Journal of Animal Sciences,
Vol.3 No.4,
October
28,
2013
ABSTRACT: The study examined the effect of human Chorionic Gonadotrophin (hCG) and/or Insulin Transferring Sodium Selenite (ITS) on the proliferation and development of rat Leydig cells purified by Nycodenz gradient. Leydig cells purity, viability and proliferation after purification and 3 days of cultured were evaluated. Leydig cells 1 × 106 cells/ml were cultured in DMEM containing 10% Newborn Calf Serum (NBCS) and divided into four kinds of treatments 1) as a control, 2) control + 2.5 IU/ml hCG, 3) control + ITS (5 μg/ml insulin, 10 μg/ml transferrin, 5 μg/ml Se), and 4) control + hCG + ITS. Leydig cells purification results showed 91.40% of purity, viability was 98.17% and concentration 7.03 × 106 cells/ml. The addition of ITS and hCG + ITS in DMEM produced Leydig cell proliferation by 88.35% and 90.64% higher than in controls (86.82%) (p p p