tm3 mouse leydig cell complete medium  (Procell Inc)

Journal: Asian Journal of Andrology

Article Title: Exosomes derived from BMSCs ameliorate cyclophosphamide-induced testosterone deficiency by enhancing the autophagy of Leydig cells via the AMPK-mTOR signaling pathway

doi: 10.4103/aja202286

Figure Lengend Snippet: BMSCs-exos ameliorate CPTD by enhancing LC autophagy in vivo . ( a ) Immunofluorescence staining of LC3 (green) in the testes of mice (scale bar = 50 μm). ( b ) Immunofluorescence staining of 3β-HSD (red) in the testes of mice (scale bar = 50 μm). ( c ) Mean fluorescence intensity of LC3 was normalized to the negative control. ( d ) Mean fluorescence intensity of 3β-HSD was normalized to the negative control. ( e ) Serum testosterone levels of mice in each group (6 mice per group). * P < 0.05, ** P < 0.01, *** P < 0.001. BMSCs: bone marrow mesenchymal stem cells; exos: exosomes; BMSCs-exos: BMSCs-derived exosomes; CP: cyclophosphamide; LCs: Leydig cells; 3β-HSD: 3β-hydroxysteroid dehydrogenase; CPTD: cyclophosphamide-induced testosterone deficiency; DAPI: 4’,6-diamidino-2-phenylindole; 3-MA: 3-methyladenine.

Article Snippet: The cells were incubated in TM3 mouse Leydig cell complete medium (Procell), and cultured at 37°C in a humidified incubator containing 5% CO 2 .

Techniques: In Vivo, Immunofluorescence, Staining, Fluorescence, Negative Control, Derivative Assay

Journal: Asian Journal of Andrology

Article Title: Exosomes derived from BMSCs ameliorate cyclophosphamide-induced testosterone deficiency by enhancing the autophagy of Leydig cells via the AMPK-mTOR signaling pathway

doi: 10.4103/aja202286

Figure Lengend Snippet: BMSCs-exos inhibit cell death in CP-exposed TM3 mouse LCs. ( a ) BMSCs-exos could be taken up by TM3 mouse LCs. ( b ) CCK-8 analysis of TM3 mouse LC viability at different concentrations of CP. ( c ) CCK-8 analysis of TM3 mouse LC viability after treatment with different concentrations of BMSCs-exos. ( d ) The Bcl-2 and Bax protein expression levels were measured with western blot. ( e ) Quantitative analysis of protein expression levels of Bcl-2 and Bax. ** P < 0.01, *** P < 0.001. BMSCs: bone marrow mesenchymal stem cells; exos: exosomes; BMSCs-exos: BMSCs-derived exosomes; CP: cyclophosphamide; LC: Leydig cell; DAPI: 4’,6-diamidino-2-phenylindole; Bcl-2: B-cell lymphoma-2; Bax: Bcl-2 associated X; NC: negative control.

Article Snippet: The cells were incubated in TM3 mouse Leydig cell complete medium (Procell), and cultured at 37°C in a humidified incubator containing 5% CO 2 .

Techniques: CCK-8 Assay, Expressing, Western Blot, Derivative Assay, Negative Control

Journal: Asian Journal of Andrology

Article Title: Exosomes derived from BMSCs ameliorate cyclophosphamide-induced testosterone deficiency by enhancing the autophagy of Leydig cells via the AMPK-mTOR signaling pathway

doi: 10.4103/aja202286

Figure Lengend Snippet: BMSCs-exos promote autophagy in CP-exposed TM3 mouse LCs. ( a ) Quantitative real-time PCR (qPCR) analysis of LC3 , Beclin-1 , and p62 mRNA expression levels in TM3 mouse LCs. ( b ) LC3-II/LC3-I, Beclin-1, and p62 protein expression levels were measured with western blot. ( c ) Quantitative analysis of protein expression levels of LC3-II/LC3-I, Beclin-1, and p62. ( d ) Immunofluorescence staining of LC3 (green) in the TM3 mouse LCs. ( e ) Mean fluorescence intensity of LC3 was normalized to the negative control. * P < 0.05, ** P < 0.01, *** P < 0.001. BMSCs: bone marrow mesenchymal stem cells; exos: exosomes; BMSCs-exos: BMSCs-derived exosomes; CP: cyclophosphamide; LCs: Leydig cells; DAPI: 4’,6-diamidino-2-phenylindole; 3-MA: 3-methyladenine; PCR: polymerase chain reaction; NC: negative control; LC3: microtubule-associated protein 1A/1B-light chain 3.

Article Snippet: The cells were incubated in TM3 mouse Leydig cell complete medium (Procell), and cultured at 37°C in a humidified incubator containing 5% CO 2 .

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Immunofluorescence, Staining, Fluorescence, Negative Control, Derivative Assay, Polymerase Chain Reaction

Journal: Asian Journal of Andrology

Article Title: Exosomes derived from BMSCs ameliorate cyclophosphamide-induced testosterone deficiency by enhancing the autophagy of Leydig cells via the AMPK-mTOR signaling pathway

doi: 10.4103/aja202286

Figure Lengend Snippet: BMSCs-exos improve the testosterone synthesis in CP-exposed TM3 mouse LCs. ( a ) Quantitative real-time PCR (qPCR) analysis of StAR and CYP11A1 mRNA expression in LCs. ( b ) The StAR and CYP11A1 protein expression levels were measured with western blot. ( c ) Quantitative analysis of protein expression levels of StAR and CYP11A1. ( d ) The testosterone concentration in TM3 mouse LC-conditioned medium was detected by ELISA. * P < 0.05, ** P < 0.01, *** P < 0.001. BMSCs: bone marrow mesenchymal stem cells; exos: exosomes; BMSCs-exos: BMSCs-derived exosomes; CP: cyclophosphamide; LCs: Leydig cells; 3-MA: 3-methyladenine; PCR: polymerase chain reaction; StAR: steroidogenic acute regulatory protein; CYP11A1: recombinant cytochrome P450 11A1; ELISA: enzyme-linked immunosorbent assay; NC: negative control.

Article Snippet: The cells were incubated in TM3 mouse Leydig cell complete medium (Procell), and cultured at 37°C in a humidified incubator containing 5% CO 2 .

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Polymerase Chain Reaction, Recombinant, Negative Control

Journal: Asian Journal of Andrology

Article Title: Exosomes derived from BMSCs ameliorate cyclophosphamide-induced testosterone deficiency by enhancing the autophagy of Leydig cells via the AMPK-mTOR signaling pathway

doi: 10.4103/aja202286

Figure Lengend Snippet: BMSCs-exos regulate AMPK-mTOR signaling pathway in CP-exposed TM3 mouse LCs. ( a ) The AMPK-mTOR signaling pathway protein levels were measured with western blot. ( b ) Quantitative analysis of protein expression levels of p-AMPK and p-mTOR. * P < 0.05, ** P < 0.01, *** P < 0.001. BMSCs: bone marrow mesenchymal stem cells; exos: exosomes; BMSCs-exos: BMSCs-derived exosomes; CP: cyclophosphamide; LCs: Leydig cells; 3-MA: 3-methyladenine; AMPK: adenosine monophosphate-activated protein kinase; p-AMPK: photophosphorylation-AMPK; mTOR: mammalian target of rapamycin; p-mTOR: photophosphorylation-mTOR; NC: negative control.

Article Snippet: The cells were incubated in TM3 mouse Leydig cell complete medium (Procell), and cultured at 37°C in a humidified incubator containing 5% CO 2 .

Techniques: Western Blot, Expressing, Derivative Assay, Negative Control