Characterization of Nestin-positive stem Leydig cells as a potential source for the treatment of testicular Leydig cell dysfunction

Abstract

The ability to identify and isolate lineage-specific stem cells from adult tissues could facilitate cell replacement therapy. Leydig cells (LCs) are the primary source of androgen in the mammalian testis, and the prospective identification of stem Leydig cells (SLCs) may offer new opportunities for treating testosterone deficiency. Here, in a transgenic mouse model expressing GFP driven by the Nestin (Nes) promoter, we observed Nes-GFP+ cells located in the testicular interstitial compartment where SLCs normally reside. We showed that these Nes-GFP+ cells expressed LIFR and PDGFR-α, but not LC lineage markers. We further observed that these cells were capable of clonogenic self-renewal and extensive proliferation in vitro and could differentiate into neural or mesenchymal cell lineages, as well as LCs, with the ability to produce testosterone, under defined conditions. Moreover, when transplanted into the testes of LC-disrupted or aging models, the Nes-GFP+ cells colonized the interstitium and partially increased testosterone production, and then accelerated meiotic and post-meiotic germ cell recovery. In addition, we further demonstrated that CD51 might be a putative cell surface marker for SLCs, similar with Nestin. Taken together, these results suggest that Nes-GFP+ cells from the testis have the characteristics of SLCs, and our study would shed new light on developing stem cell replacement therapy for testosterone deficiency.

Regulation of seminiferous tubule-associated stem Leydig cells in adult rat testes

“…PDGFRα, a cell surface protein, had been used to concentrate stem Leydig cells from neonatal (9,10) and adult (16) rats. CD51 and p75NTR also are cell surface proteins whose expressions were found to overlap with that of nestin in the neonatal mouse testis (11). By reanalysis of an earlier array study (13), we also identified a number of cell surface proteins whose mRNAs were highly expressed by stem cells but turned off with the transit of these cells to progenitor Leydig cells.…”

“…The isolation of pure stem cells requires specific cell surface markers. Several proteins had been reported in previous studies to be expressed by stem Leydig cells in neonatal or adult testes, including nestin (11,18), COUP-TFII (24,34), Arx (35), CD51 (11), p75NTR (11), and PDGFRα (9,10,16). Nestin, COUP-TFII, and Arx are intracellular proteins and, therefore, could not readily be used as markers to purify the cells.…”

“…There now is compelling evidence that most, if not all, of the adult Leydig cells arise from stem cells, not from the transdifferentiation of fetal Leydig cells (8,9). In previous studies, we and others isolated cells from neonatal testes that expressed platelet-derived growth factor receptor-α (PDGFα) or nestin (9)(10)(11). Depending on culture conditions, these cells were shown to be capable of proliferating indefinitely or of differentiating into T-producing Leydig cells and, thus, were identified as stem Leydig cells (9)(10)(11).…”

“…In previous studies, we and others isolated cells from neonatal testes that expressed platelet-derived growth factor receptor-α (PDGFα) or nestin (9)(10)(11). Depending on culture conditions, these cells were shown to be capable of proliferating indefinitely or of differentiating into T-producing Leydig cells and, thus, were identified as stem Leydig cells (9)(10)(11). During their differentiation, these cells proceeded through two intermediate stages, progenitor Leydig cells and immature Leydig cells, before becoming adult Leydig cells (12).…”

Cardiac Nestin⁺ Cells Derived from Early Stage of Dilated Cardiomyopathy Enhanced the Survival of the Doxorubicin-Injured Cardiac Muscle HL-1 Cells

“…13,14) Except for neural stem cells, Nestin also have been reported to be expressed in many other tissues, such as the bone marrow, kidney, testis, and hair follicle of the skin, which participate in cell renewal, proliferation, differentiation, and tissue regeneration as well as repair. [15][16][17][18][19] Importantly, previous studies have reported that Nestin was up-regulated in the infarcted myocardium within 48 hours, and further indicated that Nestin + cells participate in the wound healing NESTIN + CELLS ENHANCE SURVIVAL OF DOXORUBICIN INDUCED HL-1 CELLS after myocardial injury. 20,21) However, whether Nestin + cells participated in the pathogenesis of the DCM has not been reported yet.…”