Primary Cultures and Cell Lines for <i>In Vitro</i> Modeling of the Human Adrenal Cortex

Abstract

The human adrenal cortex is a complex endocrine organ that produces mineralocorticoids, glucocorticoids and androgens. These steroids are produced in distinct cell types located within the glomerulosa, fasciculata and reticularis of the adrenal cortex. Abnormal adrenal steroidogenesis leads to a variety of diseases that can cause hypertension, metabolic syndrome, infertility and premature adrenarche. The adrenal cortex can also develop steroid-producing adenomas and rarely adrenocortical carcinomas. In vitro cell culture models provide important tools to study molecular and cellular mechanisms controlling both the physiologic and pathologic conditions of the adrenal cortex. In addition, the presence of multiple steroid-metabolizing enzymes within adrenal cells makes it a model for defining possible endocrine disruptors that might block these enzymes. The regulation and dysregulation of human adrenal steroid production and cell division/tumor growth can be studied using freshly isolated cells but this requires access to human adrenal glands, which are not available to most investigators. Immortalized human adrenocortical cell lines have proven to be of considerable value in studying the molecular and biochemical mechanisms controlling adrenal steroidogenesis and tumorigenesis. Current human adrenal cell lines include the original NCI-H295 and its substrains: H295A, H295R, HAC13, HAC15, HAC50 and H295RA as well as the recently established MUC-1, CU-ACC1 and CU-ACC2. The current review will discuss the use of primary cultures of fetal and adult adrenal cells as well as adrenocortical cell lines as in vitro models for the study of human adrenal physiology and pathophysiology.

Transcriptomic Response Dynamics of Human Primary and Immortalized Adrenocortical Cells to Steroidogenic Stimuli

“…To understand the transcriptome-wide steroidogenic response of human adrenocortical cells, we performed RNA-seq on human adult adrenocortical cells stimulated with either AngII or ACTH ( Figure 1 A). Primary adult adrenocortical cells were isolated from human adrenal glands removed from kidneys marked for transplant donation [ 22 , 30 ].…”

“…Another caveat to this study is the use of primary adrenocortical cells from a single donor. Although primary cell cultures are a valuable tool in this field, there is limited access to fresh donor adrenal tissue and not all primary cells isolated from individual donors pass the rigorous screening process [ 21 , 22 , 30 ]. As such, we cannot account for variability in expression response between individuals and do not know whether the expression responses observed in these primary cells occur in all healthy adult adrenals.…”

“…To understand the transcriptome-wide steroidogenic response of human adrenocortical cells, we performed RNA-seq on human adult adrenocortical cells stimulated with either AngII or ACTH (Figure 1A). Primary adult adrenocortical cells were isolated from human adrenal glands removed from kidneys marked for transplant donation [22,30]. We characterized the timing of stimulation-induced gene expression responses by assaying five different post-stimulation time points (3,6,12,24, and 48 h), in triplicate.…”

“…There has been some effort to compare this carcinoma cell line to normal adrenal tissue, but the comparisons are largely biochemical [ 18 , 19 , 20 , 21 ]. Primary adrenocortical cells cultured from healthy tissue have also been used to study stimuli-induced adrenocortical steroidogenesis [ 22 ]. However, these cells are used less frequently because they require access to human tissue, specialized expertise in primary cell culture, and can only be maintained for a limited number of cell divisions.…”

The role of lipid second messengers in aldosterone synthesis and secretion

“…These include primary cultures derived from different species (e.g., human, bovine, murine) and a few adrenocortical carcinoma cell lines (e.g., human H295R, an H295R clone, HAC15, (17) and Y1 mouse cells). (18)(19)(20)(21)…”