Paulo Marcelo Perin

Human fallopian tube: a new source of multipotent adult mesenchymal stem cells discarded in surgical procedures.

BackgroundThe possibility of using stem cells for regenerative medicine has opened a new field of investigation. The search for sources to obtain multipotent stem cells from discarded tissues or through non-invasive procedures is of great interest. It has been shown that mesenchymal stem cells (MSCs) obtained from umbilical cords, dental pulp and adipose tissue, which are all biological discards, are able to differentiate into muscle, fat, bone and cartilage cell lineages. The aim of this study was to isolate, expand, characterize and assess the differentiation potential of MSCs from human fallopian tubes (hFTs).MethodsLineages of hFTs were expanded, had their karyotype analyzed, were characterized by flow cytometry and underwent in vitro adipogenic, chondrogenic, osteogenic, and myogenic differentiation.ResultsHere we show for the first time that hFTs, which are discarded after some gynecological procedures, are a rich additional source of MSCs, which we designated as human tube MSCs (htMSCs).ConclusionHuman tube MSCs can be easily isolated, expanded in vitro, present a mesenchymal profile and are able to differentiate into muscle, fat, cartilage and bone in vitro.

Effects of exposure to high levels of particulate air pollution during the follicular phase of the conception cycle on pregnancy outcome in couples undergoing in vitro fertilization and embryo transfer.

The objective of this retrospective cohort study was to assess the potential effects of preconceptional short-term exposure to particulate air pollution in a real-world situation on pregnancy outcome in infertile women evaluating the possible role of IVF/embryo transfer treatment on this outcome using women who had conceived naturally for the first time during the same time frame as a matched control group. The study provides evidence for an association between brief exposure to high levels of ambient particulate matter (aerodynamic diameter <or=10 microm) during the preconceptional period and early pregnancy loss, regardless of the method of conception, and showed a 2.6-fold increase in risk of miscarriage, suggesting a threshold instead of a monotonic effect of this exposure on reproductive outcome.

Human Endometrial-Derived Mesenchymal Stem Cells Suppress Inflammation in the Central Nervous System of EAE Mice

Mesenchymal stromal cells (MSCs) are undifferentiated multipotent cells endowed with the capacity for self-renewal and the potential to differentiate into several distinct cell lineages [1]. It is well established that adult MSCs constitute a reservoir found within connective tissues of most organs, and whose biological function is involved in the maintenance and repair of tissues throughout the postnatal life of an individual. Over the past years we and others have shown that menstrual blood, the endometrium and fallopian tubes are very rich sources of MSCs and able to differentiate into different cell lineages in vitro and/or in vivo [2, 3]. The unique regenerative capacity of the human endometrium following menstruation, postpartum, surgical procedures (uterine curettage, endometrial ablation) and in postmenopausal women undergoing hormonal replacement therapy suggests that MSCs niches present in this tissue are responsible, at least in part, for this process. This makes these cells a very interesting approach to studies in regenerative medicine, especially in autoimmune disorders. Multiple sclerosis (MS) is a debilitating and neurodegenerative autoimmune disease with a significant social burden. It is mainly characterized by central nervous system (CNS) inflammation, gliosis, neuronal death and demyelination [4, 5]. Its murine model, experimental autoimmune encephalomyelitis (EAE), has generated many important data concerning MS pathology and treatment [6–9]. In EAE, mice are subcutaneously immunized with myelin-derived antigens such as myelin oligodendrocyte glycoprotein (MOG35-55), myelin basic protein (MBP) and also proteolipoprotein (PLP) [6]. In the periphery T CD4 cells differentiate into Th1 and Th17 cells and have been implicated in the pathogenesis of EAE [10–12]. Although IFN-γ and ILStem Cell Rev and Rep (2012) 8:940–952 DOI 10.1007/s12015-011-9338-3

Biological effects and dose-response assessment of diesel exhaust particles on in vitro early embryo development in mice

An increased risk of early pregnancy loss in women briefly exposed to high levels of ambient particulate matter during the preconceptional period was recently observed. The effects of this exposure on early embryo development are unknown. This study was designed to assess the dose-response and biological effects of diesel exhaust particles (DEP) on in vitro embryo development using the in vitro fertilization (IVF) mouse model. Zygotes obtained from superovulated mice after IVF were randomly cultured in different DEP concentrations (0, 0.2, 2, and 20 microg/cm(2)) for 5 days and observed for their capacity to attach and develop on a fibronectin matrix until day 8. Main outcome measures included blastocyst rates 96 and 120 h after insemination, hatching discriminatory score, total cell count, proportion of cell allocation to inner cell mass (ICM) and trophectoderm (TE), ICM morphology, attachment rate and outgrowth area, apoptosis and necrosis rates, and Oct-4 and Cdx-2 expression. Multivariate analysis showed a negative dose-dependent effect on early embryo development and hatching process, blastocyst cell allocation, and ICM morphology. Although blastocyst attachment and outgrowth were not affected by DEP, a significant impairment of ICM integrity was observed in day 8 blastocysts. Cell death through apoptosis was significantly higher after DEP exposure. Oct-4 expression and the Oct-4/Cdx-2 ratio were significantly decreased in day 5 blastocysts irrespective of DEP concentration. Results suggest that DEP appear to play an important role in disrupting cell lineage segregation and ICM morphological integrity even at lower concentrations, compromising future growth and viability of the blastocyst.

Comparison of the efficacy of two commercially available media for culturing one-cell embryos in the in vitro fertilization mouse model

OBJECTIVE To examine the effects of two commercial media on the development of mouse ova fertilized in vitro to the blastocyst stage. DESIGN Animal model. SETTING Academic institution. ANIMAL(S) Eight-week old, superovulated mice. INTERVENTION(S) One-cell embryos cultured in vitro up to the blastocyst stage in potassium-enriched simplex optimized medium (KSOM) or G1/G2 medium. MAIN OUTCOME MEASURE(S) Blastocyst and hatching rates, total cell number count, and proportion of allocation of cells to the inner cell mass (ICM) and trophectoderm (TE). RESULT(S) The percentage of zygotes that developed to the blastocyst stage 96 and 120 hours after insemination was statistically significantly higher in the KSOM group. The percentage of blastocysts that partially or completely hatched by day 5 of culture was 84% and 71% for the KSOM and G1/G2 groups, respectively, showing a statistically significant difference between the groups. The mean number of ICM cells was 11.7 +/- 4.0 and 9.2 +/- 5.2 for the zygotes cultured in KSOM and G1/G2 media, respectively, revealing a statistically significantly higher cell number in the ICM of blastocysts derived from culture in KSOM medium. The ICM/TE ratio in the blastocysts cultured in KSOM or G1/G2 media was similar in both groups. CONCLUSION(S) Commercially available KSOM medium is superior to sequential G1/G2 media for culturing one-cell embryos up to the blastocyst stage in the mouse IVF model.

In vitro fertilization, embryo development, and cell lineage segregation after pre- and/or postnatal exposure of female mice to ambient fine particulate matter

OBJECTIVE To evaluate effects of pre- and/or postnatal exposure to ambient fine particulate matter on fertilization, embryo development, and cell lineage segregation in preimplantation blastocysts using the IVF mouse model. DESIGN Animal model. SETTING Academic institution. ANIMAL(S) Six-week-old, superovulated mice. INTERVENTION(S) Pre- and postnatal exposure to filtered air (FA-FA), filtered-ambient air (FA-AA), or ambient air (AA-AA) in exposure chambers 24 hours a day for 9 weeks. MAIN OUTCOME MEASURE(S) Gestation length, litter size, sex ratio, ovarian response to superovulation, fertilization rate, embryo development, blastocyst and hatching rates, total cell count, and proportion of cell allocation to inner-cell mass (ICM) and trophectoderm (TE). RESULT(S) Gestation length, litter size and birth weight, live-birth index, and sex ratio were similar among exposure groups. Ovarian response was not affected by the exposure protocol. A multivariate effect for pre- and/or postnatal exposure to ambient fine particulate matter on IVF, embryo development, and blastocyst differential staining was found. Cell counts in ICM and ICM/TE ratios in blastocysts produced in the FA-FA protocol were significantly higher than in blastocysts produced in the FA-AA and AA-AA protocols. No difference in total cell count was observed among groups. CONCLUSION(S) Our study suggests that exposure to ambient fine particulate matter may negatively affect female reproductive health by disrupting the lineage specification at the blastocyst stage without interfering in early development of the mouse embryo.

Human Tubal-Derived Mesenchymal Stromal Cells Associated with Low Level Laser Therapy Significantly Reduces Cigarette Smoke–Induced COPD in C57BL/6 mice

Cigarette smoke-induced chronic obstructive pulmonary disease is a very debilitating disease, with a very high prevalence worldwide, which results in a expressive economic and social burden. Therefore, new therapeutic approaches to treat these patients are of unquestionable relevance. The use of mesenchymal stromal cells (MSCs) is an innovative and yet accessible approach for pulmonary acute and chronic diseases, mainly due to its important immunoregulatory, anti-fibrogenic, anti-apoptotic and pro-angiogenic. Besides, the use of adjuvant therapies, whose aim is to boost or synergize with their function should be tested. Low level laser (LLL) therapy is a relatively new and promising approach, with very low cost, no invasiveness and no side effects. Here, we aimed to study the effectiveness of human tube derived MSCs (htMSCs) cell therapy associated with a 30mW/3J—660 nm LLL irradiation in experimental cigarette smoke-induced chronic obstructive pulmonary disease. Thus, C57BL/6 mice were exposed to cigarette smoke for 75 days (twice a day) and all experiments were performed on day 76. Experimental groups receive htMSCS either intraperitoneally or intranasally and/or LLL irradiation either alone or in association. We show that co-therapy greatly reduces lung inflammation, lowering the cellular infiltrate and pro-inflammatory cytokine secretion (IL-1β, IL-6, TNF-α and KC), which were followed by decreased mucus production, collagen accumulation and tissue damage. These findings seemed to be secondary to the reduction of both NF-κB and NF-AT activation in lung tissues with a concomitant increase in IL-10. In summary, our data suggests that the concomitant use of MSCs + LLLT may be a promising therapeutic approach for lung inflammatory diseases as COPD.

The effect of recombinant human luteinizing hormone on oocyte/embryo quality and treatment outcome in down-regulated women undergoing in vitro fertilization

Objective: To compare oocyte/embryo quality and treatment outcome in patients undergoing IVF/ICSI using recombinant luteinizing hormone (rhLH) or human menopausal gonadotrophin (hMG) as a supplement to recombinant follicle-stimulating hormone (rhFSH) during ovarian stimulation. Methods: After pituitary desensitization, sixty women undergoing their first IVF treatment cycle were randomized to receive a fixed daily dose of either rhFSH (225 IU/d) plus hMG (75 IU/d) (group A, n=30) or rhLH (75 IU/d) (group B, n=30). The number/quality of metaphase II (MII) oocytes, zygote/embryo quality, and clinical pregnancy/implantation rates were compared between the two groups. Results: The mean number of normal/abnormal MII oocytes was 4.5±3.10/2.8±3.13 in group A and 2.4±1.47/4.0±3.70 in group B, showing a significant difference between groups (p=0.006; p=0.01; respectively). The fertilization rate was similar in both groups, yet the number of zygotes judged normal was significantly higher (p= 0.02) in group A (2.4±2.40) as compared to group B (1.2±1.76). The mean number and quality score of embryos at transfer were similar in both groups. Overall clinical pregnancy/implantation rates were 46.4%/25.3% in group A and 38.1%/17.1% in group B. The trend toward better pregnancy outcomes among patients in group A did not reach statistical significance. Conclusions: Our study suggests that the addition of recombinant luteinizing hormone instead of human menopausal gonadotrophin to recombinant follicle-stimulating hormone throughout ovulation induction in down-regulated women undergoing IVF does not improve ovarian response and has a negative impact on oocyte/zygote quality. The result is a trend toward poorer treatment outcome.