It seems that PRP has a protective effect on ovarian failure in the infertile female rat model.

Cleavage-stage embryos have an absolute requirement for pyruvate and lactate, but as the morula compacts, it switches to glucose as the preferred energy source to fuel glycolysis. Substrates such as glucose, amino acids, and lactate are moved into and out of cells by facilitated diffusion. In the case of lactate and pyruvate, this occurs via H 1 -monocarboxylate cotransporter (MCT) proteins. To clarify the role of MCT in development, transport characteristics for DL-lactate were examined, as were mRNA expression and protein localisation for MCT1 and MCT3, using confocal laser scanning immunofluorescence in freshly collected and cultured embryos. Blastocysts demonstrated significantly higher affinity for DL-lactate than zygotes (K m 20 6 10 vs 87 6 35 mmol lactate/l; P 5 0.03 by linear regression) but was similar for all stages. For embryos derived in vivo and those cultured with glucose, MCT1 mRNA was present throughout preimplantation development, protein immunoreactivity appearing diffuse throughout the cytoplasm with brightest intensity in the outer cortical region of blastomeres. In expanding blastocysts, MCT1 became more prominent in the cytoplasmic cortex of blastomeres, with brightest intensity in the polar trophectoderm. Without glucose, MCT1 mRNA was not expressed, and immunoreactivity dramatically reduced in intensity as morulae died. MCT3 mRNA and immunoreactivity were not detected in early embryos. The differential expression of MCT1 in the presence or absence of glucose demonstrates that it is important in the critical regulation of pH and monocarboxylate transport during preimplantation development, and implies a role for glucose in the control of MCT1, but not MCT3, expression.

neurons like other living cells may have ultraweak photon emission (Upe) during neuronal activity. this study is aimed to evaluate Upe from neural stem cells (nSc) during their serial passaging and differentiation. We also investigate whether the addition of silver nanoparticles (AgNPs) or enhancement of UPE (by AgNPs or mirror) affect the differentiation of NSC. In our method, neural stem and progenitor cells of subventricular zone (SVZ) are isolated and expanded using the neurosphere assay. the obtained dissociated cells allocated and cultivated into three groups: groups: i: cell (control), ii: cell + mirror, and iii: cell + Agnps. After seven days, the primary neurospheres were counted and their mean number was obtained. Serial passages continuous up to sixth passages in the control group. Differentiation capacity of the resulting neurospheres were evaluated in vitro by immunocytochemistry techniques. Measurement of Upe was carried out by photomultiplier tube (pMt) in the following steps: at the end of primary culture, six serial cell passages of the control group, before and after of the differentiation for 5 minutes. The results show that neither mirror nor AgNPs affect on the neurosphere number. The UPE of the NSC in the sixth subculturing passage was significantly higher than in the primary passage (P < 0.05). AgNPs significantly increased the UPE of the NSC compared to the control group before and after the differentiation (P < 0.05). Also, the treatment with AgNPs increased 44% neuronal differentiation of the harvested NSCs. UPE of NSC after the differentiation was significantly lower than that before the differentiation in each groups, which is in appropriate to the cell numbers (P < 0.0001). The mirror did not significantly increase UPE, neither before nor after the differentiation of nSc. As a conclusion, nSc have Upe-properties and the intensity is increased by serial passaging that are significant in the sixth passage. The AgNPs increases the UPE intensity of NSC that pushes more differentiation of NSC to the neurons. The mirror was not effective in enhancement of UPE. As a result, UPE measurement may be suitable for assessing and studying the effects of nanoparticles in living cells and neurons.

Bone morphogenetic protein 4 (BMP4) and retinoic acid (RA) signaling are the key regulators for germ cell and meiosis induction, respectively. Gonadal tissue also provides an appropriate microenvironment for oocyte differentiation in vivo. The current study aimed to determine whether mimicking in vivo niche is more efficient for oocyte differentiation from embryonic stem (ES) cells. Here, differentiation of mouse ES cells toward oocyte-like cells using embryoid body (EB) and monolayer protocols was induced in the presence (+BMP4) or absence (-BMP4) of BMP4. On day 5, each group was co-cultured with ovarian somatic cells in the presence or absence of RA (+RA or -RA) for an additional 14 days. Our results showed a significant increase in expression of meiotic markers in the +BMP4 condition in EB differentiation protocol. Further differentiation with ovarian somatic cells led to a subpopulation of oocyte-like cell formation. Compared to the controls, the +RA condition resulted in a significant elevation of the meiotic gene expression in contrast to Oct4 that significantly decreased in both protocols. In the cells pre-treated with BMP4 and then exposed to RA in the monolayer differentiation protocol, the gene expression levels of germ cell, Mvh, and maturation markers, Cx37, Zp2, and Gdf9, were also upregulated significantly. Therefore, it can be concluded that +BMP4 and +RA along with ovarian somatic cell co-culture improved the rate of in vitro oocyte differentiation.

IntroductionNeural stem cells (NSCs) reside along the ventricular axis of the mammalian brain. They divide infrequently to maintain themselves and the down‐stream progenitors. Due to the quiescent property of NSCs, attempts to deplete these cells using antimitotic agents such as cytosine b‐Aarabinofuranoside (Ara‐C) have not been successful. We hypothesized that implementing infusion gaps in Ara‐C kill paradigms would recruit the quiescent NSCs and subsequently eliminate them from their niches in the subventricular zone (SVZ).MethodsWe infused the right lateral ventricle of adult mice brain with 2% Ara‐C using four different paradigms—1: one week; 2: two weeks; 3, 4: two weeks with an infusion gap of 6 and 12 h on day 7. Neurosphere assay (NSA), neural colony‐forming cell assay (N‐CFCA) and immunofluorescent staining were used to assess depletion of NSCs from the SVZ.ResultsNeurosphere formation dramatically decreased in all paradigms immediately after Ara‐C infusion. Reduction in neurosphere formation was more pronounced in the 3rd and 4th paradigms. Interestingly 1 week after Ara‐C infusion, neurosphere formation recovered toward control values implying the presence of NSCs in the harvested SVZ tissue. Unexpectedly, N‐CFCA in the 3rd paradigm, as one of the most effective paradigms, did not result in formation of NSC‐derived colonies (colonies >2 mm) even from SVZs harvested 1 week after completion of Ara‐C infusion. However, formation of big colonies with serial passaging capability, again confirmed the presence of NSCs.ConclusionsOverall, these data suggest Ara‐C kill paradigms with infusion gaps deplete NSCs in the SVZ more efficiently but the niches would repopulate even after the most vigorous kill paradigm used in this study.