녹용 유래 중간엽 줄기세포의 체외 배양 효율 증진

Abstract

The annual regrowth of deer antlers is a connatural developmental event in mammals. Therefore, studying regeneration of deer antlers could be a unique natural model of rapid and complete bone regeneration in human and other mammals. However, little is known about culture conditions and regulatory factors that stimulate growing of deer antler cells in vitro. The aim of this study was to enhance an in vitro culture efficiency of mesenchymal stem cells (MSCs) derived from deer antlers. In order to improve the culture condition, we selected minimal essential medium alpha (MEM alpha) as a basal medium and investigate whether serum could stimulate growing in these cells in basal medium in a dose-dependent manner. Next, to investigate the optimal temperature and O-2 tension, the antler cells were cultured in different temperature and controlled O-2 percentages. Through the results of number of harvested cells after 1 week, we selected MEM alpha, 10% fetal bovine serum (FBS), 37 degrees C, 20% O-2, and 5% CO2 tension as a basic culture conditions. Also, we could observed enhanced proliferation results by addition of the supplements [L-glutamine 2 mM, beta-mercaptoethanol 100 mu M, non-essential amino acid (NEAA) 0.1 mM, and HEPES 10 mM] and growth factors [basic fibroblast growth factor (bFGF) 10 ng/mL, epidermal growth factor (EGF) 20 ng/mL, insulin-like growth factor-1 (IGF-1) 10 ng/mL] and harvested antler cells strongly expressed STRO-1 and CD 90. Our results demonstrate that allow continuous proliferation of antler cells in vitro established the foundation to basic biology of antler cells and makes possible application to the regenerative medicine in a broad sence.