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

Jiang MH, Cai B, Tuo Y, Wang J, Zang ZJ, Tu X, Gao Y, Su Z, Li W, Li G, Zhang M, Jiao J, Wan Z, Deng C, Lahn BT, Xiang AP - Cell Res. (2014)

Bottom Line: 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.In addition, we further demonstrated that CD51 might be a putative cell surface marker for SLCs, similar with Nestin.

View Article: PubMed Central - PubMed

Affiliation: 1] Cell-gene Therapy Translational Medicine Research Center, The Third Affiliated Hospital, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong 510630, China [2] Key Laboratory for Stem Cells and Tissue Engineering, Center for Stem Cell Biology and Tissue Engineering, Ministry of Education, Sun Yat-sen University, Guangzhou, Guangdong 510080, China [3] Department of Anatomy, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong 510080, China.

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.

Show MeSH

Related in: MedlinePlus

Nes-GFP+ cells differentiated into LCs and facilitated testosterone production in the testes of 22-month-old mice after transplantation. (A) Immunofluorescence staining showed the accumulation of cells positive for PKH26 (red) and P450scc (green) or LHR (green) in the interstitial area of testes of old mice 10 days after Nes-GFP+ cell implantation. The expression levels of P450scc and LHR in the 22-month-old mice were lower than those in the young mice (3-month-old). Scale bar, 50 μm. (B) In the 22-month-old mice, the serum testosterone concentrations were significantly increased in the Nes-GFP+ cell-treated mice compared to the saline-treated old mice (*P < 0.05, **P < 0.01). n = 6. Young/Saline (+), 3-month-old mice receiving saline injection; Old/Saline (+), 22-month-old mice receiving saline injection; Old/Cells (+), 22-month-old mice receiving Nes-GFP+ cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4260348&req=5

fig8: Nes-GFP+ cells differentiated into LCs and facilitated testosterone production in the testes of 22-month-old mice after transplantation. (A) Immunofluorescence staining showed the accumulation of cells positive for PKH26 (red) and P450scc (green) or LHR (green) in the interstitial area of testes of old mice 10 days after Nes-GFP+ cell implantation. The expression levels of P450scc and LHR in the 22-month-old mice were lower than those in the young mice (3-month-old). Scale bar, 50 μm. (B) In the 22-month-old mice, the serum testosterone concentrations were significantly increased in the Nes-GFP+ cell-treated mice compared to the saline-treated old mice (*P < 0.05, **P < 0.01). n = 6. Young/Saline (+), 3-month-old mice receiving saline injection; Old/Saline (+), 22-month-old mice receiving saline injection; Old/Cells (+), 22-month-old mice receiving Nes-GFP+ cells.

Mentions: In male mice, aging generally goes along with a progressive decline in the biosynthesis of testosterone by LCs24,25,26. To test whether Nes-GFP+ cells could restore testosterone production in a senescence model, we injected PKH26-labeled Nes-GFP+ cells into the testes of 22-month-old mice. Without cell transplantation, the expression levels of P450scc and LHR in the LCs of these old mice were dramatically decreased (Figure 8A), and these mice also exhibited a lower serum testosterone level compared with young adults injected with saline (Figure 8B). Ten days after cell transplantation, the PKH26-labeled Nes-GFP+ cells colonized the interstitium of the testis and strongly expressed P450scc and LHR (Figure 8A). Notably, the serum testosterone levels of mice with cell transplantation were increased significantly compared with the control mice with saline injection (Figure 8B). Taken together, these results demonstrate that the Nes-GFP+ cells possess key properties of SLCs as they are capable of colonizing the interstitium and differentiating into mature testosterone-producing LCs.


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

Jiang MH, Cai B, Tuo Y, Wang J, Zang ZJ, Tu X, Gao Y, Su Z, Li W, Li G, Zhang M, Jiao J, Wan Z, Deng C, Lahn BT, Xiang AP - Cell Res. (2014)

Nes-GFP+ cells differentiated into LCs and facilitated testosterone production in the testes of 22-month-old mice after transplantation. (A) Immunofluorescence staining showed the accumulation of cells positive for PKH26 (red) and P450scc (green) or LHR (green) in the interstitial area of testes of old mice 10 days after Nes-GFP+ cell implantation. The expression levels of P450scc and LHR in the 22-month-old mice were lower than those in the young mice (3-month-old). Scale bar, 50 μm. (B) In the 22-month-old mice, the serum testosterone concentrations were significantly increased in the Nes-GFP+ cell-treated mice compared to the saline-treated old mice (*P < 0.05, **P < 0.01). n = 6. Young/Saline (+), 3-month-old mice receiving saline injection; Old/Saline (+), 22-month-old mice receiving saline injection; Old/Cells (+), 22-month-old mice receiving Nes-GFP+ cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4260348&req=5

fig8: Nes-GFP+ cells differentiated into LCs and facilitated testosterone production in the testes of 22-month-old mice after transplantation. (A) Immunofluorescence staining showed the accumulation of cells positive for PKH26 (red) and P450scc (green) or LHR (green) in the interstitial area of testes of old mice 10 days after Nes-GFP+ cell implantation. The expression levels of P450scc and LHR in the 22-month-old mice were lower than those in the young mice (3-month-old). Scale bar, 50 μm. (B) In the 22-month-old mice, the serum testosterone concentrations were significantly increased in the Nes-GFP+ cell-treated mice compared to the saline-treated old mice (*P < 0.05, **P < 0.01). n = 6. Young/Saline (+), 3-month-old mice receiving saline injection; Old/Saline (+), 22-month-old mice receiving saline injection; Old/Cells (+), 22-month-old mice receiving Nes-GFP+ cells.
Mentions: In male mice, aging generally goes along with a progressive decline in the biosynthesis of testosterone by LCs24,25,26. To test whether Nes-GFP+ cells could restore testosterone production in a senescence model, we injected PKH26-labeled Nes-GFP+ cells into the testes of 22-month-old mice. Without cell transplantation, the expression levels of P450scc and LHR in the LCs of these old mice were dramatically decreased (Figure 8A), and these mice also exhibited a lower serum testosterone level compared with young adults injected with saline (Figure 8B). Ten days after cell transplantation, the PKH26-labeled Nes-GFP+ cells colonized the interstitium of the testis and strongly expressed P450scc and LHR (Figure 8A). Notably, the serum testosterone levels of mice with cell transplantation were increased significantly compared with the control mice with saline injection (Figure 8B). Taken together, these results demonstrate that the Nes-GFP+ cells possess key properties of SLCs as they are capable of colonizing the interstitium and differentiating into mature testosterone-producing LCs.

Bottom Line: 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.In addition, we further demonstrated that CD51 might be a putative cell surface marker for SLCs, similar with Nestin.

View Article: PubMed Central - PubMed

Affiliation: 1] Cell-gene Therapy Translational Medicine Research Center, The Third Affiliated Hospital, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong 510630, China [2] Key Laboratory for Stem Cells and Tissue Engineering, Center for Stem Cell Biology and Tissue Engineering, Ministry of Education, Sun Yat-sen University, Guangzhou, Guangdong 510080, China [3] Department of Anatomy, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong 510080, China.

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.

Show MeSH
Related in: MedlinePlus