Generation of induced pluripotent stem cells (iPSCs) IRMBi002-A from an Alzheimer's disease patient carrying a D694N mutation in the APP gene.

Induced pluripotent stem cells (iPSC) were generated from skin fibroblasts obtained from a 58 year-old woman suffering from Alzheimer's disease and carrying a D694N mutation on Amyloid precursor protein (APP). Fibroblasts were reprogrammed into iPSC using the integration-free Sendai Virus which allows the expression of the Yamanaka factors. Verification of their pluripotency was achieved by demonstrating the expression of pluripotency markers and their differentiation potential into the three primary germ layers. The cells have the corresponding mutation and present a normal karyotype. The reported APP-D694N iPSC line may be used to model and study human AD pathology in vitro.

Generation of induced pluripotent stem cells (IRMBi001-A) from an Alzheimer's disease patient carrying a G217D mutation in the PSEN1 gene.

Induced pluripotent stem cells (iPSC) were generated from skin fibroblasts obtained from a 50 year-old patient suffering from Alzheimer's disease and carrying a G217D causal mutation on presenilin 1 (PSEN1). iPSCs were obtained following reprogramming using the integration-free Sendai Virus system which allows expression of the Yamanaka factors. Verification of their pluripotency was achieved by demonstrating the expression of pluripotency markers and their differentiation potential into the three primary germ layers. iPS cells carry the patient G217D mutation and present a normal karyotype. The reported PS1-G217D iPSC line may be used to model and study human AD pathology in vitro.

In-vitro processing of sperm with autoantibodies and in-vitro fertilization results.

In-vitro fertilization and embryo transfer were carried out in twenty infertile couples with significant levels of anti-sperm antibodies on the ejaculated spermatozoa. Over 21 cycles (15 couples), 95 mature oocytes were collected and inseminated with spermatozoa obtained by swim-up migration after rapid dilution and washing of the ejaculates. An overall fertilization rate of 38.9% was obtained with these post-migration (PM) preparations. When greater than 70% of the inseminated spermatozoa were covered with both IgG and IgA antibodies, only 14% of the 43 oocytes were fertilized. A higher fertilization rate was obtained with the PM preparations containing less than 70% of spermatozoa coated with one or both classes of antibodies. Under these conditions 60% of the 52 oocytes were fertilized. Fertility rate correlated better with IgG than with IgA antibody levels. In order to decrease the proportion of antibody-coated spermatozoa in the inseminated populations, washed spermatozoa were immuno-adsorbed on Mage's plates before swim-up migration. Over 11 cycles (10 couples) 52 mature oocytes were inseminated with these post-migration immuno-depleted sperm preparations (PMP) containing less than 65% of antibody-coated spermatozoa: 31% of the oocytes were fertilized. This rate compares favourably to IVF results obtained with the PM preparations with greater than 70% of spermatozoa coated with both classes of antibodies. In five couples who had different IVF attempts with the two sperm preparations, the immuno-depletion resulted in a slight increase in the fertilization rates: 10% for the PM preparations versus 26% for the PMP preparations. Sperm binding to the zona pellucida was decreased in the majority of unsuccessful attempts.(ABSTRACT TRUNCATED AT 250 WORDS)

[In vivo and in vitro fertilizing ability of immature human epididymal spermatozoa]. / Fécondance in vivo et in vitro de spermatozoïdes épididymaires humains immatures.

In cases of congenital absence of vas deferens (9 patients) or after failure of previous epididymovasostomy (2 patients), in vitro fertilization (IVF) was attempted with spermatozoa surgically obtained at the epididymal caput level. These sperm populations showed little progressive motility (5.9 +/- 6.5%) and an marked necrozoospermia (19.3 +/- 17.4%). Stimulation by caffeine (4.5 mM) alone or associated with heterologue normal seminal fluid resulted in most of the cases in an initiation of motility with an improvement of the progressive velocity. In 9 IVF attempts, 31 mature oocytes were inseminated with 5.10(3) to 1.5.10(6) motile spermatozoa. The dynamic characteristics in 3 inseminated sperm populations were Vp (24.2 +/- 8.3 microns/s), Ah (8.6 +/- 2.0 microns) at room temperature. Sperm binding to zona pellucida was decreased (0 to about 20 spermatozoa per oocyte) and there was no fertilization. In the same period, 21 attempts of intra uterine insemination and 14 attempts of intracervical inseminations were made in 5 couples who remained infertile after patent high epididymovasostomy (4) or vasovasostomy (1) and having immature spermatozoa stimulated as previously described. Antisperm antibodies were detected on the ejaculated spermatozoa in four men. No pregnancy was obtained with these immature stimulated spermatozoa. The fertility of the female partners was confirmed in 3 women after insemination with donor sperm.