Abstract
The rhesus monkey embryonic stem cell line R366.4 has been identified to differentiate into a number of cell types. However, it has not been well characterized for its response to drugs affecting reproductive endocrinology. Kisspeptins (KPs) are ligands for the GPR-54, which is known to modulate reproductive function. The current study was designed to determine the effect of the KP-10 peptide on R366.4 cells and to investigate the role of KP-GPR54 in the cell proliferation process. Four different doses (0.1, 1, 10, and 100 nM) of KP-10 and control were selected to evaluate cell growth parameters and cellular morphological changes over a 72 hr period. The cells were treated with kisspeptin-10 during the early rosette stage. Proliferation rates, analyzed by flow cytometry and cell count methods, were found to be decreased after treatment. Moreover, the number of rosettes was found to decrease following KP-10 treatments. Morphological changes consisting of neuronal projections were also witnessed. This suggested that KP-10 had an antiproliferative effect on R366.4 cells leading to a differentiation state and morphological changes consistent with neuronal stem cell development. The R366.4 stem cell line differentiates based on kisspeptin signaling and may be used to investigate reproductive cell endocrinology in vitro.
Highlights
Embryonic stem cells (ESC) are capable of self-renewal and differentiation into cells of the three germ cell layers [1,2,3,4]
One of the great advantages of rhesus monkey ESCs is that they share many characteristics with human ESCs, including cellular morphology, surface marker expression, and developmental potential [10], as well as greater than 90% DNA homology [27,28,29,30], which allows for better mimicry of the human system
The growth and differentiation of the R366.4 stem cell line is modulated by reproductive endocrine kisspeptin
Summary
Embryonic stem cells (ESC) are capable of self-renewal and differentiation into cells of the three germ cell layers [1,2,3,4]. 26 ESC lines from the rhesus macaque, an animal often used as a model to study human disease, have been produced from in vivo flushed blastocysts [7, 9]. Among these are eight R series cell lines (4 males and 4 females) made from blastocysts produced in vivo by Dr James Thomson at the Wisconsin National Primate Research Center [6]. The R366.4 cell line has not been well characterized for physiological changes in response to drug treatments
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