Summary:We compared UCB mononuclear cells (MNC) with CD34؉ selected cells in a serum-free static culture system. Cell number proliferation of MNCs was inferior to CD34 ؉ selected cells. MNCs, however, showed a substantial increase from 0.94% CD34 ؉ cells on day 0 to 5.8% on day 7, whereas in the CD34؉ selected samples the CD34؉ cell content declined continously from 62.2% on day 0 to 27.7% on day 7. The number of CFU-GM increased during culture of both cell fractions. Here, only the MNCs showed a substantial increase in clonogenicity on day 7 and day 14 to 11.1-and 4.1-fold input, respectively. This expansion of the CD34 ؉ progenitor cell pool in the MNCs fraction was at least in part attributable to T cells, since the physical abrogation of T cells blocked this effect. Refeeding and reseeding of cells on day 7 had stimulating effects especially on the CD34؉ cells, where cell number proliferation increased from 16.3-fold without to 58.1-fold on day 14. Also, we could find sporadic chromosomal aberrations in four of 100 metaphases examined after 7-20 days of ex vivo expansion. The significance of this observation needs to be clarified in a larger series.
Background:Physical activity has been shown to stimulate haematopoiesis in patients with anaemia due to chronic renal failure or haematological malignancies.Objective:To evaluate the effect of moderate exercise on the production of haematopoietically active factors.Methods:Ten patients (four men and six women, mean (SD) age 51 (10) years) with a haemoglobin concentration under 130 g/l (men) or 120 g/l (women) carried out five three minute exercise bouts at an intensity of 80% of the maximal heart rate, corresponding to a lactate concentration of 3 (0.5) mmol/l. Patients rested for three minutes between bouts. The concentrations of interleukin 6, stem cell factor, granulocyte-monocyte colony stimulating factor, granulocyte colony stimulating factor, erythropoietin, and growth hormone (GH) were evaluated before and in the eight hours after exercise.Results:GH had risen significantly 15 minutes after exercise (1.1 (1.3)v2.7 (2.8) ng/ml; p<0.05). No change in the concentration of the other cytokines and growth factors was observed in the eight hours after exercise.Conclusions:In patients with anaemia, submaximal exercise does not affect the concentration of haematopoietically active cytokines. However, it leads to an increased concentration of GH. This may be responsible for the improved haematopoiesis observed after an exercise programme in patients with chronic diseases.
Recombinant human granulocyte colony-stimulating factor (rhG-CSF) mobilized peripheral blood progenitor cells (PBPCs) from healthy individuals are a rapidly emerging alternative source to bone marrow for allogeneic transplantation. Although widely applied in the meantime, only limited information on feasibility and safety of mobilization and collection of PBPCs is currently available from prospective multicenter studies specifically designed to investigate this donation modality. This ongoing multicenter study on the performance as well as the short- and long-term safety profile of rhG-CSF-induced mobilization and collection of PBPCs was initiated in October 1999. The study is designed to recruit a total of 300 healthy family donors who will be followed regularly for a period of 5 years after donation. The first interim report presented here summarizes results obtained after enrollment of 150 donors from nine German institutions. The study protocol allowed the individual choice between two dose regimens of rh-CSF (10 micro g/kg per day vs 2x8 micro g/kg per day of donor body weight). The primary endpoint was defined as a yield of > or =5x10(6) CD34(+) cells/kg of recipient body weight in a single leukapheresis product. This endpoint was attained by 50% of donors receiving the lower rhG-CSF dose regimen and by 75% of donors with the higher dose regimen ( p<0.0009). A total of 478 acute adverse events attributable to the mobilization procedure were recorded and manifested predominantly as transient bone pain and headaches (80%). No persistent hematologic or nonhematologic adverse events have occurred in this study so far. Thus, the current experience in a prospective multicenter study supports previous single-center and retrospective registry reports in that the collection of PBPCs after rhG-CSF mobilization is feasible and associated with frequent, but generally mild and acceptable side effects.
Summary:all only 30% of patients in need of an allogeneic transplantation have a suitable family donor. 1 The search for an unrelated donor is expensive, time-consuming and transplanOne of the main limiting factors for increased use of human umbilical cord blood (UCB) in adult allogeneic tation is associated with an increased risk of graft rejection, GVHD and opportunistic infections. 2 UCB as an alternative transplantation is the small number of progenitor cells that can be collected and infused. Ex vivo expansion of source of hematopoietic stem cells can be easily collected, cryopreserved and stored. Of great interest is the reported UCB might help to overcome this limitation. Whether an expansion of UCB cells will also lead to co-expansion low rate of acute grade III-IV GVHD after UCB transplantation. [3][4][5] Moreover, chronic GVHD seems to be uncomof contaminating maternal cells, and thus may alter graft characteristics and lead to an increased incidence mon. 5,6 One of the main limiting factors for UCB transplantation, however, is the limited volume of UCB that can of GVHD, has not been looked at so far. We initiated cultures with UCB mononuclear cells (MNC) in a stanbe collected and thus the relatively small number of total nucleated cells and hematopoietic progenitor cells that can dard medium containing stem cell factor (SCF), flt-3L, Il-3, IL-6, EPO and G-CSF. To address the question of be infused. 5 Therefore, most experience with UCB transplantation has been obtained in the pediatric setting. To contaminating maternal cells we performed interphase FISH analysis of the X and Y chromosome simulincrease the number of hematopoietic progenitor cells and to make UCB transplantation suitable for the adult patient, taneously. Male (XY) cord blood samples were investigated for maternal (XX) cells at day 0 and at several ex vivo expansion of UCB is gaining more and more attention. time points during culture. We could not detect maternal cells in any of the nine samples studied whenWith the first reported UCB transplantation in 1988 for a patient with Fanconi anemia 7 concern has been raised cultures were started at day 0. Culturing did not expand previously undetected maternal cells into a range that about the presence of maternal cells in cord blood which might lead to possible vertical transmission of infectious could be seen with FISH technology, as all samples remained negative for maternal cells throughout culture agents and severe GVHD. Since maternal cells may only share one HLA haplotype with the recipient, a substantial periods of 14 days. We then artificially contaminated male UCB with maternal mononuclear cells at concen-HLA incompatibility may be present. 8 Several groups investigating the presence of maternal cells reported contrations of 5 and 15% at day 0. After 14 days, maternal MNC were still detectable, but the percentage was flicting results. [9][10][11][12][13][14][15][16] To our knowledge, the behavior of contaminating maternal cells during in vitro expansion of reduced to 1.7% and 6%, re...
Androgens stimulate development and growth of the external male genitalia. Since hypospadias represents the most common congenital abnormality in the male newborn and the mechanism of action in this disorder is still unclear, androgen binding was assessed in cultured fibroblasts from biopsies from genital skin of 10 patients with idiopathic hypospadias. For comparison, binding was determined in corresponding samples from 8 males with normal penile development and from 9 patients with known androgen resistance syndromes (testicular feminization, Reifenstein syndrome, pseudovaginal perineoscrotal hypospadias). Finally, binding was measured in 10 samples of nongenital skin. Maximum specific binding (Bmax) in idiopathic hypospadias varied from 3.2 to 15.5 (median 6.6) fmol.mg protein-1. Bmax in samples of persons with normal genital development was between 12.2 and 17.9 fmol.mg protein-1 (median 13.2). Bmax in samples of patients with known androgen resistance syndromes was exactly in the range reported previously in the literature. It is evident that Bmax in samples of patients with idiopathic hypospadias differs significantly (P less than 0.01), (Mann Whitney U-test) from those with normal genital development. Thus it seems reasonable to conclude that in some patients with idiopathic hypospadias the genital defect is caused by receptor deficiency.
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