Effect of icariin on the transformation efficiency of induced pluripotent stem cells into sperm cells in vitro / Estudio de los efectos de icariina en la eficiencia de la transformación de espermatozoides extrovertidos inducidos en cuerpos multicompetentes

Objective: To investigate the effect of icariin on the transformation efficiency of germ cell-like cells from mouse induced pluripotent stem cells into sperm cells in vitro. Methods: Firstly, mouse induced pluripotent stem cells were induced and cultured to transform into germ cell-like cells, and the primordial germ cell-like cells were identified by Western blot and RT-PCR. Then, different concentrations of icariin (0.1μg/mL, 1μg/mL, 10μg/mL and 100μg/mL) were added into the culture medium, and the obtained primitive germ cell-like cells were cultured, Western blot and RT-PCR were used to identify the obtained sperm cells, the transformation efficiency was compared. Results: The primordium germ cell-like cells obtained from mouse induced pluripotent stem cells in vitro specially expressed Oct-4 protein, C-kit protein, Mvh mRNA, Fragilis mRNA and Stella mRNA. The sperm cells were specially expressed VASA, SCP3 and γH2AX proteins. RT-PCR showed that the sperm cells were specially expressed Ddx4, Tp2 and Prm1 mRNA. Compared with the control group, the expression level of VASA protein (1.744±0.283, 2.882±0.373, 6.489±0.460), SCP3 protein (2.250±0.306, 7.058±0.521, 8.654±0.804), γH2AX protein (4.304±0.433, 5.713±0.339, 9.268±0.545), Ddx4 mRNA (1.374±0.145, 2.846±0.194, 4.021±0.154), Tp2 mRNA (1.358±0.130, 3.623±0.326, 5.811±0.390) and Prm1 mRNA (1.326±0.162, 3.487±0.237, 4.666±0.307) in 0.1μg/mL, 1μg/mL, 10μg/mL icariin experimental groups were all lower than that of VASA protein (10.560±0.413), SCP3 protein (13.804±0.642), γH2AX protein (11.874±0.464), Ddx4 mRNA (6.4005±0.361), Tp2 mRNA (7.314±0.256) and Prm1 mRNA (7.334±0.390) in 100μg/mL icariin experimental group. (AU)

Objetivo: Investigar el efecto de icariina en la eficiencia de la conversión in vitro inducida en espermatozoides de cultivos de células germinativas derivadas de la transformación de células madre pluripotentes inducidas de ratón. Métodos: Primero se indujeron y cultivaron células madre pluripotentes inducidas de ratón para transformarlas en células similares a las células germinales, y las células similares a las células germinales primordiales se identificaron mediante Western blot y RT-PCR. A continuación, se añadieron diferentes concentraciones de icariina (0,1μg/mL, 1μg/mL, 10μg/mL and 100μg/mL) al medio de cultivo, y se cultivaron las células primitivas similares a células germinales obtenidas, se utilizaron Western blot y RT-PCR para identificar las células espermáticas obtenidas, y se comparó la eficacia de la transformación. Resultados: Las células germinales primitivas obtenidas in vitro a partir de células madre pluripotentes inducidas de ratón expresaron especialmente la proteína Oct-4, la proteína C-kit, el ARNm de Mvh, el ARNm de Fragilis y el ARNm de Stella. Los espermatozoides expresaban especialmente las proteínas VASA, SCP3 y γH2AX. La RT-PCR mostró que los espermatozoides expresaban especialmente los ARNm Ddx4, Tp2 y Prm1. En comparación con el grupo de control, el nivel de expresión de la proteína VASA (1,744±0,283; 2,882±0,373; 6,489±0,460), la proteína SCP3 (2,250±0,306; 7,058± 0,521; 8,654±0,804), proteína γH2AX (4,304±0,433; 5,713±0,339; 9,268±0,545), ARNm Ddx4 (1,374±0,145; 2,846±0,194; 4,021±0,154), ARNm Tp2 (1,358±0,130; 3,623±0,326; 5,811±0,390) y ARNm Prm1 (1,326±0,162; 3,487±0,237; 4,666±0,307) en grupos experimentales de 0,1μg/mL, 1μg/mL, 10μg/mL de icariina fueron todos más bajos que los de la proteína VASA (10,560±0,413), proteína SCP3 (13,804±0,642), proteína γH2AX (11,874±0,464), ARNm Ddx4 (6,4005±0,361), ARNm Tp2 (7,314±0,256) y ARNm Prm1 (7,334±0,390) en 100μg/mL icariina grupo experimental. (AU)

Effect of icariin on the transformation efficiency of induced pluripotent stem cells into sperm cells in vitro.

OBJECTIVE: To investigate the effect of icariin on the transformation efficiency of germ cell-like cells from mouse induced pluripotent stem cells into sperm cells in vitro. METHODS: Firstly, mouse induced pluripotent stem cells were induced and cultured to transform into germ cell-like cells, and the primordial germ cell-like cells were identified by Western blot and RT-PCR. Then, different concentrations of icariin (0.1µg/mL, 1µg/mL, 10µg/mL and 100µg/mL) were added into the culture medium, and the obtained primitive germ cell-like cells were cultured, Western blot and RT-PCR were used to identify the obtained sperm cells, the transformation efficiency was compared. RESULTS: The primordium germ cell-like cells obtained from mouse induced pluripotent stem cells in vitro specially expressed Oct-4 protein, C-kit protein, Mvh mRNA, Fragilis mRNA and Stella mRNA. The sperm cells were specially expressed VASA, SCP3 and γH2AX proteins. RT-PCR showed that the sperm cells were specially expressed Ddx4, Tp2 and Prm1 mRNA. Compared with the control group, the expression level of VASA protein (1.744±0.283, 2.882±0.373, 6.489±0.460), SCP3 protein (2.250±0.306, 7.058±0.521, 8.654±0.804), γH2AX protein (4.304±0.433, 5.713±0.339, 9.268±0.545), Ddx4 mRNA (1.374±0.145, 2.846±0.194, 4.021±0.154), Tp2 mRNA (1.358±0.130, 3.623±0.326, 5.811±0.390) and Prm1 mRNA (1.326±0.162, 3.487±0.237, 4.666±0.307) in 0.1µg/mL, 1µg/mL, 10µg/mL icariin experimental groups were all lower than that of VASA protein (10.560±0.413), SCP3 protein (13.804±0.642), γH2AX protein (11.874±0.464), Ddx4 mRNA (6.4005±0.361), Tp2 mRNA (7.314±0.256) and Prm1 mRNA (7.334±0.390) in 100µg/mL icariin experimental group. CONCLUSIONS: Icariin can promote the transformation of mouse induced pluripotent stem cells into sperm cells in vitro, and it is concentration-dependent manner in a certain concentration range.

Effect of advanced glycosylation end products (AGEs) on proliferation of human bone marrow mesenchymal stem cells (MSCs) in vitro.

This study aims to explore the effect of advanced glycosylation end products (AGEs) on proliferation of human bone marrow mesenchymal stem cells in vitro and the underlying mechanism. Bone marrow cell proliferation was determined by WST-8 assay using Cell Counting Kit-8 under the intervention of AGEs. In addition, the content of maldondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were also measured. The proliferation activity of mesenchymal stem cells (MSCs) was significantly inhibited when AGEs were added to culture medium, and this effect was dose-dependent and time-dependent. As the concentration of AGEs-bovine serum albumin increased, the content of intracellular MDA was significantly increased, but the activity of SOD in cell homogenates was significantly suppressed, which also showed a dose-dependent manner. AGEs could significantly inhibit the proliferation of MSCs in vitro by improving the oxidative stress in MSCs and breaking the homeostasis of intracellular environment.

[Diagnosis and treatment of ovotesticular disorders of sex development in children].

OBJECTIVE: To investigate the diagnosis and treatment of ovotesticular disorders of sex development (DSD) in children. METHODS: We reviewed the clinical data of 9 cases of ovotesticular DSD admitted in our department from 1988 to 2007. RESULTS: The patients ranged in age from 9 months to 9 years, 7 raised as males and 2 as females. As for the karyotype, 4 cases were 46,XX, 2 were 46,XX/46,XY, 1 was 46,XY, and the other 2 had no karyotype data. All of them presented with obscure external genitalia: perineal or penoscrotal hypospadias with or without cryptorchidism in males and hypertrophy of the clitoris in females. They were diagnosed with ovotesticular DSD by gonad biopsy and underwent genitoplasty. CONCLUSION: The gender assignment of the ovotesticular DSD patient was chiefly based on the development of external genitalia, dominant gonad, karyotype and the parent's will. Laparoscopic technology is recommended in gonad biopsy and orchiopexy during the treatment of ovotesticular DSD.