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Biological Analysis of Leydig Cells-Conditioned Medium To Support Rat Bone Marrow Mesenchymal Stem Cells Differentiation

ANNALES BOGORIENSES Vol 22, No 1 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

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Abstract

The developed Leydig cells-conditioned medium (LCM) contains bioactive materials secreted by Leydig cells in vitro.  LCM was used to evaluate the ability of bone marrow mesenchymal stem cells differentiation. Bone marrow mesenchymal stem cells (1x 106 cell/ml) were cultured in : 1) DMEM supplemented with 10% NBCS as a control (M), 2) M supplemented with 10 ng/ml testosterone; 3) M supplemented with 50%  LCM ; 4) M supplemented with 50% LCM and 2.5 IU/ml hCG. Bone marrow mesenchymal stem cells that were cultured with  LCM has a positive reaction (57.4%) to histochemistry staining 3β-HSD and produced 1.87 ng/ml testosterone. Supplementation of hCG to LCM  increased the positive number of Leydig cells and testosterone production by 74.6% and 12.33 ng/ml (P<0.05). It can be concluded that Leydig cells-conditioned medium can support differentiation of bone marrow mesenchymal stem cells into Leydig cells.

Maternal Contribution In Revealing The Effects Of Methoxyacetic Acid (Maa) Administered Before Implantation On The Embryonic Development Of Swiss Webster Mice (Mus musculus)

ANNALES BOGORIENSES Vol 9, No 1 (2004): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

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Abstract

Maternal contribution to and direct action of methoxyacetic acid (MAA) on the embryonic development bad been examined by conducting embryo tTansfer. To reveal the maternal contribut ion, compacted morulae and early blastocysts, which were collected from untreated Swiss Webster donor mice on day 3 of gestation, were transferred to day 2 pseudopregnant recipients, after having been treated with 2 .0 mmollkg body weight (b.w.) MAA by gavage on day 1 of pseudopregnancy. Direct effect of MAA on the embryonic development were observed by transferring compacted morulae and early bla tocysts, simi larly recovered from day 3 pregnant donor mice, after MAA treatment on day 2 of gestation with the same method and dosing, to untreated day 2 pseudopregnant recipients. Control donor mice and recipient were given distilled water only as the MAA olvent. Observations on fetuses resulting fTom embryo transfer wert: carried out on day 16 of gestatlOn . Administration of MAA to the donors tended to decrease the unplantatlOn rate and the survival rate of the implanted embryos. W11en MAA was given to the recipients the implantation rate and survival rate of embryos transferred decreased significantly (p<0 .05) but the survival rate of implanted embryos were significantly higher (p<0.05) Lf compared to those of MAA treated donors. The intrauterine death tended to inc rease either in the treated donors or recipients. Th re was no et1ect of MAA on the fetal body weight and in producing fetal malformations . It is concluded that at the beginning of implantation, maternal contribution in revealing the effects of MAA on the embryonic development of Swiss Webster mice is predominant, whereas after Implantation took place, the quality of the embryos become ;lore important for their survival.

INFLUENCES OF INCUBATION TIME AND SUCROSE CONCENTRATION ON MICE (Mus musculus L.) OOCYTE VIABILITY FOR ENUCLEATING PROCEDURE

Jurnal Kedokteran Hewan Vol 12, No 3 (2018): J. Ked. Hewan
Publisher : Syiah Kuala University

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Abstract

This study aimed to find out the optimum incubation time to complete mouse oocyte maturation at Metaphase II (MII) stage and determine the optimum sucrose concentration enabling to induce nuclear swelling for visualization that is important for enucleating process at the initial procedure of somatic cell nuclear transfer (SCNT). In this current study, mice were used as animal model. Completely randomized design was arranged, consists of 2 trials with 4 treatments and 7 replications. In the first trial, the oocytes were cultured at 0-2, 4-6, 8-10, and 12-14 h in 5% CO2 incubator at 37 C. Second, the MII oocytes obtained from previous trial were cultured in M199 medium containing different concentrations of sucrose (0, 1.5, 3, and 6%). The parameters measured were the oocyte viability at various stages, i.e germinal vesicle (GV), metaphase I (MI), anaphase/telophase I (A/T I), and metaphase II (MII), and the viability of swollen nuclear oocytes using Hoechst/PI staining. The results showed that the optimum incubation time required by oocytes to reach MII stage was 12-14 h with a percentage of 57.14±12.67%, while the optimum sucrose concentration for nuclear swelling was found at 3% with a percentage of 100±0.00%. Our findings provided preliminary results related to the maturation process of the mouse oocyte nucleus, which is meaningful for the initial procedure of SCNT.