Effect of human ovarian tissue vitrification/warming on the expression of genes related to folliculogenesis

Ovarian tissue cryopreservation is an alternative strategy to preserve the fertility of women predicted to undergo premature ovarian failure. This study was designed to evaluate the expression of folliculogenesis-related genes, including factor in the germline alpha [FIGLA], growth differentiation factor-9 [GDF-9], follicle-stimulating hormone receptor [FSHR], and KIT LIGAND after vitrification/warming of human ovarian tissue. Human ovarian tissue samples were collected from five transsexual women. In the laboratory, the ovarian medullary part was removed by a surgical blade, and the cortical tissue was cut into small pieces. Some pieces were vitrified and warmed and the others were considered as non-vitrified group [control]. Follicular normality was assessed with morphological observation by a light microscope, and the expression of FIGLA, KIT LIGAND, GDF- 9, and FSHR genes was examined using real-time RT-PCR in both the vitrified and non-vitrified groups. Overall, 85% of the follicles preserved their normal morphologic feature after warming. The percentage of normal follicles and the expression of FIGLA, KIT LIGAND, GDF-9, and FSHR genes were similar in both vitrified and non-vitrified groups [P > 0.05]. Vitrification/warming of human ovarian tissue had no remarkable effect on the expression of folliculogenesis-related genes

Histone modification of embryonic stem cells produced by somatic cell nuclear transfer and fertilized blastocysts

Nuclear transfer-embryonic stem cells [NT-ESCs] are genetically identical to the donor's cells; provide a renewable source of tissue for replacement, and therefore, decrease the risk of immune rejection. Trichostatin A [TSA] as a histone deacetylase inhibitor [HDACi] plays an important role in the reorganization of the genome and epigenetic changes. In this study, we examined whether TSA treatment after somatic cell nuclear transfer [SCNT] can improve the developmental rate of embryos and establishment rate of NT-ESCs line, as well as whether TSA treatment can improve histone modification in NT-ESCs lines. In this experimental study, mature oocytes were recovered from BDF1 [C57BL/6xDBA/2] mice and enucleated by micromanipulator. Cumulus cells were injected into enucleated oocytes as donor. Reconstructed embryos were activated in the presence or absence of TSA and cultured for 5 days. Blastocysts were transferred on inactive mouse embryonic fibroblasts [MEF], so ESCs lines were established. ESCs markers were evaluated by reverse transcription-polymerase chain reaction [RT-PCR]. Histone modifications were analyzed by enzyme linked immunosorbent assay [ELISA]. Result of this study showed that TSA treatment after SCNT can improve developmental rate of embryos [21.12 +/- 3.56 vs.8.08 +/- 7.92], as well as establishment rate of NT-ESCs line [25 vs.12.5]. We established 6 NT-ESCs in two experimental groups, and three embryonic stem cells [ESCs] lines as control group. TSA treatment has no effect in H3K4 acetylation and H3K9 tri-methylation in ESCs. TSA plays a key role in the developmental rate of embryos, establishment rate of ESC lines after SCNT, and regulation of histone modification in NT-ESCs, in a manner similar to that of ESCs established from normal blastocysts

Mesenchymal stem cells as an alternative for schwann cells in rat spinal cord injury

Spinal cord has a limited capacity to repair; therefore, medical interventions are necessary for treatment of injuries. Transplantation of Schwann cells has shown a great promising result for spinal cord injury [SCI]. However, harvesting Schwann cell has been limited due to donor morbidity and limited expansion capacity. Furthermore, accessible sources such as bone marrow stem cells have drawn attentions to themselves. Therefore, this study was designed to evaluate the effect of bone marrow-derived Schwann cell on functional recovery in adult rats after injury. Mesenchymal stem cells were cultured from adult rats' bone marrow and induced into Schwann cells in vitro. Differentiation was confirmed by immunocytochemistry and RT-PCR. Next, Schwann cells were seeded into collagen scaffolds and engrafted in 3 mm lateral hemisection defects. For 8 weeks, motor and sensory improvements were assessed by open field locomotor scale, narrow beam, and tail flick tests. Afterwards, lesioned spinal cord was evaluated by conventional histology and immunohistochemistry. In vitro observations showed that differentiated cells had Schwann cell morphology and markers. In this study, we had four groups [n = 10 each]: laminectomy, control, scaffold and scaffold + Schwann cells. Locomotor and sensory scores of cell grafted group were significantly better than control and scaffold groups. In histology, axonal regeneration and remyelination were better than control and scaffold groups. This study demonstrates that bone marrow-derived Schwann cells can be considered as a cell source for Schwann cells in SCI treatment