Factors controlling embryonic heart cell proliferation in serum-free synthetic media |
| |
Authors: | A C Nag M Ingland M Cheng |
| |
Institution: | (1) Department of Biological Sciences, Oakland University, 48063 Rochester, Michigan |
| |
Abstract: | Summary Embryonic chick cardiac cell cultures, plated on collagen-coated dishes, containing serum-free synthetic media proliferate
actively. The basic medium contained Ham's F12 nutrient mixture, fetuin, ascorbic acid, and bovine serum albumin. This medium
was supplemented with various combinations of factors; endothelial cell growth supplement (ECGS), epidermal growth factor
(EGF), insulin (I), transferrin (T), selenium (S), hydrocortisone, and thyroxine or supplemented alone. Basic medium supplemented
with ECGS alone contributes to the highest final cell density among all other factors used in various combinations or alone.
The final cell density of the control culture with 2% fetal bovine serum was higher than those of all experimental cultures
and an additional control culture grown in the basic medium. Combinations of factors without ECGS do not promote significant
cell proliferation. Thyroxine is required to induce optimal differentiation and contractility of cardiac myocytes in vitro.
Fibronectin and laminin did not show any more influence than collagen did on the growth and maintenance of cardiac myocytes
in serum-free media. The proportion of cardiac muscle cells in ECGS-containing media was higher than those in other experimental
media and control media with the exception of ECGS and ITS-containing medium that showed lower proportion of cardiac myocytes
than that of serum-containing medium on Days 3 and 5. The profiles of incorporation of 3H]thymidine into DNA of heart cells in experimental and control cultures showed a peak in incorporation values within the
first week of culture and subsequently declined. Autoradiography studies revealed that cardiac myocytes in culture supplemented
with ECGS alone attained a peak in labeling index on Day 1 with approximately 62% labeled cells. Subsequently, the labeling
indices declined. Cardiac myocytes grown in media without ECGS showed significantly lower labeling indices than those in ECGS-containing
media. This study has demonstrated the influence of ECGS, EGF and ITS in promoting the growth of cardiac myocytes and also
in contributing to the maintenance of contractile cardiac myocytes in serum-free, long-term culture. The influence of ECGS
on heart cell proliferation is considered to be superior to that of EGF and ITS.
This study was supported in part by a grant HL-25482 from the National Heart Lung and Blood Institute and a grant from the
American Heart Association of Michigan. |
| |
Keywords: | cardiac myocytes nonmuscle cells culture proliferation growth factors |
本文献已被 SpringerLink 等数据库收录! |
|