Serum-free cultures of murine adrenal cortical cells.
OBJECTIVE
The feasibility of culturing murine adrenal cortical cells before transplantation was investigated in this study.


METHODS
Primary murine adrenal cortical cells were maintained in either fetal bovine and horse sera-containing media or serum-free media. Real-time polymerase chain reaction was used to quantify the levels of adrenal cortical gene expression in the cultured cells.


RESULTS
The use of sera-containing media led to the growth of many cells in the culture, but the expression of Sf-1, Dax-1, and Cyp11b1 in such cultures declined rapidly. In contrast, there was no significant cell growth in the serum-free culture medium. Culturing murine adrenal cortical cells in the serum-free medium resulted in higher levels of Sf-1, Dax-1, and Cyp11b1 gene expression. In the serum-free medium, adrenal cortical cells also responded to adrenocorticotropic hormone by increasing the expression of Cyp11b1 and suppressing the expression of Dax-1 in a dose-dependent manner. The addition of basic fibroblast growth factor to the serum-free medium maintained the expression of Sf-1, Dax-1, and Cyp11b1 for 4 weeks.


CONCLUSIONS
Adrenal cortical cells isolated from adult mice were successfully maintained in a serum-free culture medium with basic fibroblast growth factor. This culture system may be suitable for further manipulation of adrenal cortical cells in vitro before transplantation.
Chu
Yinting
Y
Department of Bioengineering, Henry Samueli School of Engineering, University of California, Los Angeles, CA 90095, USA.
Wu
Benjamin M
BM
McCabe
Edward R
ER
Dunn
James C Y
JC
D000302:Adrenal Cortex / Q000166:cytology*; D000818:Animals; D018929:Cell Culture Techniques; D017690:Cell Transplantation; D002478:Cells, Cultured; D057137:DAX-1 Orphan Nuclear Receptor; D004268:DNA-Binding Proteins / Q000235:genetics; D005240:Feasibility Studies; D016222:Fibroblast Growth Factor 2; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D044967:Serum; D013252:Steroid 11-beta-Hydroxylase / Q000235:genetics; D054339:Steroidogenic Factor 1; D014157:Transcription Factors / Q000235:genetics
D057137:DAX-1 Orphan Nuclear Receptor; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; C539966:Nr0b1 protein, mouse; D018160:Receptors, Cytoplasmic and Nuclear; D054339:Steroidogenic Factor 1; D014157:Transcription Factors; D016222:Fibroblast Growth Factor 2; D013252:Steroid 11-beta-Hydroxylase