Transplantation of Adrenal Cortical Progenitor Cells Enriched by Nile Red

Abstract

Background The adrenal cortex may contain progenitor cells useful for tissue regeneration. Currently there are no established methods to isolate these cells. Material and Methods Murine adrenal cells were sorted into a Nile-Red-bright (NRbright) and a Nile-Red-dim (NRdim) population of cells according to their degree of cholesterol content revealed by Nile Red fluorescence. The cells were transplanted under the renal capsule to determine their ability for regeneration. Results The NRbright cells contained an abundance of lipid droplets, whereas the NRdim cells contained little. The NRbright cells expressed Sf1 and the more differentiated adrenal cortical genes including Cyp11a1, Cyp11b1, and Cyp11b2, whereas the NRdim cells expressed Sf1 but not the more differentiated adrenal cortical genes. After 56 days of implantation in unilateral adrenalectomized mice, the NRdim cells expressed Sf1 and the more differentiated adrenal cortical genes, whereas the NRbright cells ceased to express Sf1 as well as the more differentiated adrenal cortical genes. NRdim cells also proliferated in the presence of basic fibroblast growth factor. Conclusions The population of NRdim cells contained adrenal cortical progenitor cells that can proliferate and give rise to differentiated daughter cells. These cells may be useful for adrenal cortical regeneration.

Label-Free Enrichment of Adrenal Cortical Progenitor Cells Using Inertial Microfluidics

“…However, the expression of zonal-specific genes, Cyp11b1 and Cyp11b2, were substantially suppressed for the cells collected from the outermost outlets, indicating that those cells were least differentiated (P<0.05). Collectively, the results show that more differentiated multicellular clusters of adrenal cells, expressing higher levels of zonal-specific genes (Cyp11b1 and Cyp11b2) were enriched at inner outlets (outlet 2 and 3), while the expression levels of those genes were significantly lower for the adrenal cortical progenitor cells collected at the outermost outlets (outlet 1) [25]. The clear differences in gene expression profiles of cells collected at different outlets suggest that differences in differentiation of adrenal cortical cells can be reflected in biophysical property dissimilarities (e.g., adhesive properties and relative tendency to remain in clusters after tissue dissociation).…”

Adrenocortical cell transplantation reverses a murine model of adrenal failure

“…The technique of adrenal cell transplantation in mice was described previously; 5,11 however, it had not been demonstrated that these cells could restore the adrenal function. In addition to improved survival, mice that received a sufficient dose of adrenal cells also had near normal levels of corticosterone 14 days after staged bilateral adrenalectomy.…”

“…After removing the surrounding fat, they were incubated in the digestion mixture at 37°C for 1 hour with gentle shaking. 11 The digestion mixture consisted of 10 mL of HBSS containing 2 mg/mL collagenase I, 0.05 mg/mL DNase I, and 5 mg/mL bovine serum albumin. After dispersing the cells through a pipette, they were washed with Adrenal Media and were filtered through a 70-μm strainer (Millipore, Bedford, MA).…”

“…10 We previously showed that adrenocortical cells implanted in mice continued to express adrenal-specific genes for up to 8 weeks. 5, 11 In this report, we sought to determine whether the implanted adrenocortical cells could support the function of the adrenal glands in mice. Adrenal cells harvested from adult mice were seeded onto a collagen sponge and were implanted into mice that underwent either simultaneous or staged bilateral adrenalectomy.…”

The adrenal gland microenvironment in health, disease and during regeneration

“…Considering these drawbacks related to adrenal hormone supplementation, several alternative therapeutic approaches have been proposed and tested. 129 These include auto-and allotransplantations of the adrenal glands, 130 adenoviral-associated virus (AAV)-mediated gene targeting, 131,132 and either allo-or xenotransplan-tation of adrenocortical cells, [133][134][135][136][137][138][139] or various stem cells reprogrammed into steroidogenic-like cells. [140][141][142] Apart from allotransplantation of a whole human adrenal with its intact microenvironment, all the other experiments were performed in animals, preferentially using rodent models.…”