Purification of germline stem cells from adult mammalian ovaries: a step closer towards control of the female biological clock?

For decades it was believed that a non-renewable pool of oocyte-containing follicles is established in female mammals at birth. This cornerstone of reproductive biology was challenged 5 years ago by a study reporting on the presence of mitotically-active germ cells in juvenile and adult mouse ovaries. Additional findings presented in this study and others that followed further suggested that mammals retain the capacity to generate oocytes during adulthood; however, isolation of oocyte-producing germline stem cells (GSC) as unequivocal proof of their existence remained elusive. This piece of information now appears to have been provided by Ji Wu and colleagues. In addition to showing that proliferative germ cells resembling male spermatogonial stem cells can be purified from neonatal or adult mouse ovaries and maintained in vitro for months, transplantation studies demonstrated that these cells generate oocytes in ovaries of chemotherapy-sterilized recipients that fertilize and produce viable offspring. Although these findings do not establish that oogenesis occurs in adult females under physiological conditions, they strongly support the existence of GSC in adult mouse ovaries. If equivalent cells can be found in human ovaries, stem cell-based rejuvenation of the oocyte reserve in ovaries on the verge of failure may one day be realized

Purification of germline stem cells from adult mammalian ovaries: a step closer towards control of the female biological clock?

For decades it was believed that a non-renewable pool of oocyte-containing follicles is established in female mammals at birth. This cornerstone of reproductive biology was challenged 5 years ago by a study reporting on the presence of mitotically-active germ cells in juvenile and adult mouse ovaries. Additional findings presented in this study and others that followed further suggested that mammals retain the capacity to generate oocytes during adulthood; however, isolation of oocyte-producing germline stem cells (GSC) as unequivocal proof of their existence remained elusive. This piece of information now appears to have been provided by Ji Wu and colleagues. In addition to showing that proliferative germ cells resembling male spermatogonial stem cells can be purified from neonatal or adult mouse ovaries and maintained in vitro for months, transplantation studies demonstrated that these cells generate oocytes in ovaries of chemotherapy-sterilized recipients that fertilize and produce viable offspring. Although these findings do not establish that oogenesis occurs in adult females under physiological conditions, they strongly support the existence of GSC in adult mouse ovaries. If equivalent cells can be found in human ovaries, stem cell-based rejuvenation of the oocyte reserve in ovaries on the verge of failure may one day be realized

Bone Marrow Transplantation Restores Follicular Maturation and Steroid Hormones Production in a Mouse Model for Primary Ovarian Failure

Recent studies suggest that bone marrow stem cells (BMSCs) are promising grafts to treat a variety of diseases, including reproductive dysfunction. Primary ovarian failure is characterized by amenorrhea and infertility in a normal karyotype female, with an elevated serum level of follicle-stimulating hormone (FSH) and a decrease level of estrogen caused by a mutation in FSH receptor (FSHR) gene. Currently, there is no effective treatment for this condition. The phenotype of FSHR (−/−) mouse, FORKO (follitropin receptor knockout), is a suitable model to study ovarian failure in humans. Female FORKO mice have elevated FSH, decreased estrogen levels, are sterile because of the absence of folliculogenesis, and display thin uteri and small nonfunctional ovaries. In this study, we determined the effects of BMSC transplantation on reproductive physiology in this animal model. Twenty four hours post BMSC transplantation, treated animals showed detectable estroidogeneic changes in daily vaginal smear. Significant increase in total body weight and reproductive organs was observed in treated animals. Hemotoxylin and eosin (H&E) evaluation of the ovaries demonstrated significant increase in both the maturation and the total number of the follicles in treated animals. The FSH dropped to 40–50% and estrogen increased 4–5.5 times in the serum of treated animals compared to controls. The FSHR mRNA was detected in the ovaries of treated animals. Our results show that intravenously injected BMSCs were able to reach the ovaries of FORKO mice, differentiate and express FHSR gene, make FSHR responsive to FSH, resume estrogen hormone production, and restore folliculogenesis

Bone Marrow Contribution to Renal Repair Following Ischemic Injury

Repair of the mammalian renal tubular epithelium following ischemia/reperfusion injury is largely effected by surviving epithelial cells and other cells resident in the kidney interstitium; however, the precise nature of the contribution of circulating extrarenal cells is unknown. The purpose of this study is to determine whether bone marrow cells have the capacity to differentiate into renal tubular epithelium. This was investigated using both an ex vivo transplantation model as well as an attempt to grow epithelial cells from bone marrow in a liquid culture system. Recipient female C57BL/6 mice were myeloablated via irradiation and transplanted in-travenously with lineage-depleted syngeneic male bone marrow. Following this, renal ischemia was induced by surgical interruption of the renal vascular pedicle. Thin sections were assayed for donor contribution by examination by fluorescence in situ hybridization (FISH) for the Y-chromosome. Tubules incorporating donor (Y-chromosome positive) cells were only rarely found (\u3c 2%) in the outer medulla of damaged kidneys; far more frequent were donor-derived interstitial cells, which appear not to be entirely inflammatory in phenotype. For the tissue culture experiments, collagenase-released marrow stromal cells (CR-MSC) were obtained by collagenase treatment of bone chips following mechanical dissociation of bones harvested from C57BL/6 mice. These cells were grown in culture and examined via immu-nostaining and reverse-transcription polymerase chain reaction for epithelial markers. Unfortu-nately, the resultant cells, although appearing to form epithelial colonies, were found to not ex-press the epithelial markers cytokeratin and ZO-1 by RT-PCR and immunostaining; instead they appeared to be fibroblasts with a novel morphology. In sum, bone marrow contribution to the renal epithelium appears to be a rare phenomenon at best and as of now cannot be replicated in vitro

Location of Intra- and Extracellular M. tuberculosis Populations in Lungs of Mice and Guinea Pigs during Disease Progression and after Drug Treatment

The lengthy treatment regimen for tuberculosis is necessary to eradicate a small sub-population of M. tuberculosis that persists in certain host locations under drug pressure. Limited information is available on persisting bacilli and their location within the lung during disease progression and after drug treatment. Here we provide a comprehensive histopathological and microscopic evaluation to elucidate the location of bacterial populations in animal models for TB drug development

The genetics of potential albendazole and ivermectin resistance in lymphatic filariae /

A current initiative to eliminate lymphatic filariasis (LF), headed by the World Health Organization, aims to interrupt transmission of the disease through yearly community-wide treatment with the broad spectrum anthelmintic albendazole (ABZ), in combination with ivermectin (IVM) or diethylcarbamazine (DEC). Over the years, the use of both ABZ and IVM in the treatment of veterinary parasites has led to widespread anthelmintic resistance against these drugs. In this study, we genotyped microfilaria of Wuchereria bancrofti, a causative agent of LF, in order to detect the presence of mutations which confer ABZ resistance in other parasites, and we identified such mutations in worms obtained from untreated patients in Ghana and Burkina Faso, West Africa. Microfilaria from patients who had been treated with ABZ + IVM, had a significantly higher frequency of the resistant genotype, and this frequency was even higher in worms from patients that had received two rounds of treatment. In addition, the untreated population of microfilaria had an excess of homozygotes in the population. This excess homozygosity was equivalent to a Wright's Inbreeding Statistic of FIT= 0.44, and we found that the population was significantly subdivided between patients. In order to better understand the mechanisms and factors involved in the potential spread of ABZ resistance, caused by such mutations, through a population of Culex-transmitted W. bancrofti, we developed a deterministic model that incorporates genotype structure into the epidemiological model EPIFIL. This model predicts that the combination of ABZ + DEC leads to stronger selection for the resistant genotype than ABZ + IVM, and that drug efficacy assumptions are an important factor affecting the spread of drug resistance. Treatment coverage, non-random mating, initial allele frequency and number of treatments also had substantial impact on the speed and magnitude of the spread of ABZ resistance. When we expanded this model to include potential IVM-resistance alleles we found that, under ABZ + IVM treatment, selection for resistance to either drug is enhanced by the presence of resistance against the second drug. Similarly, excess homozygosity caused by parasite non-random mating may increase selection for a dominant IVM resistance allele through enhancing the spread of a recessive ABZ resistance allele. Resistence developed more slowly when it was inherited as a polygenic trait. Results from this study suggest that resistance monitoring is crucial, as resistance may not be apparent until treatment is stopped, recrudescence occurs and treatment is reapplied

Transplantation of a line of RFP-transgenic SMSCs into chemotherapy-sterilized recipient mice.

<p>A) Immunofluorescence of ovarian sections from sterilized mice 7 days after transplantation with RFP-transgenic SMSCs. Arrows indicate RFP staining in the ovarian stroma. B) RFP staining was observed around the antral follicle 21 days after transplantation. Asterisks indicate the antral follicle. C) RFP-positive cells were observed around granulosa cells in recipient ovaries 28 days after SMSC transplantation. D) RFP staining was observed in antral follicles of recipient ovaries 2 months after SMSC transplantation. Scale bars: A, C) 50 µm; B, D) 100 µm; A, insets) 20 µm; and B, D insets) 10 µm.</p