To date, hematopoietic stem and progenitor cells from human umbilical cord blood (CB) have been employed in approximately 90 allogeneic (56 sibling and 34 unrelated) matched and mismatched transplantations worldwide with easy and successful restoration of hematopoiesis. Requests for stem cell preparations from CB will continue to increase. Thus, as a pilot study, the examination and standardization of unrelated cord blood-derived stem cell preparations and banking as well as their biologic characterization were initiated. Up to October 1995, a total of 574 samples [mean volume 79 +/- 26 ml, total nucleated cells (NC) 8.5 +/- 5 x 10(8), BFU-E 9.5 +/- 8.6 x 10(5), CFU-GM 5.7 +/- 6.3 x 10(5), CFU-GEMM 1.6 +/- 1.9 x 10(5)] from cord-derived or placental-derived residual blood have been defined by hematologic, immunologic, and microbiologic criteria. These CB samples were collected from the umbilical cord vein immediately after vaginal full-term delivery (n = 450) or cesarean section (n = 124) and stored frozen in liquid nitrogen. Seven percent of all samples collected could not be considered for potential transplants because of volumes < 40 ml. Only 5.0 ml of a CB sample is required for routine laboratory testing, consisting of HLA class I typing, HLA class II typing by sequence-specific oligonucleotide probes (PCR-SSOP), ABO typing, sterility control, assessment of progenitor and stem cells by colony-forming and LTC-IC assays, and CD34+ status. To assess the potential problem of contaminating maternal cells, a PCR was performed on 7 representative samples. During the initial 6 months of the unrelated CB collection program, a median bacterial contamination rate of 18% (20% skin flora species, 80% perineal flora species) was encountered, which has since been reduced to < 1% through practical experience. With regard to viral infections, maternal sera was tested for HBsAg (0.6% positive), anti-HCV (0%), anti-HAV (IgG 18%, IgM 0%), anti-HIV-1-2 (0%), anti-EBV (IgG 98%, IgM 0%), anti-HTLVI-II (0%), anti-CMV (IgG 43%, IgM 0.4%), toxoplasmosis (46%) and syphilis (0%). In addition, all cord blood samples were tested by PCR for CMV infection. With regard to its clinical relevance, it is important that only 0.3% of all the samples were positive for CMV by this sensitive method. This may represent a critical advantage of CB grafts over bone marrow (BM) since, in contrast, > 40% of the unrelated BM donors have been identified to be positive for CMV. An additional advantage of CB is that since 20% of CB samples were collected from ethnic minorities, it appears possible to balance common HLA types and uncommon HLA types represented in this group. In summary, with the extensive practical experience of the obstetric collection team as well as the stem cell-processing laboratory, it appears feasible to obtain a 90% yield of unrelated CB-derived stem cell preparations for banking, which clearly should meet the medical and regulatory qualification criteria required for clinical transplantation. To test the feasibility of hematopoietic ...

Summary:Cord blood (CB) is an alternative source of marrow repopulating cells for the treatment of pediatric patients. Over the past 7 years, the results of CB transplantation for pediatric Assuming a threshold of 2 × 10 7 nucleated cells (NC)/kg body weight required for transplantation and 10 ± 5 × 10 8 diseases have been promising. Over 500 unrelated and related cord blood transplants have been performed. + CB cells were cultured with the four cytokine combinations in H5100 medium, all combi-10 ± 5 × 10 8 nucleated cells (NC) stored in the CB Bank Düsseldorf, 100%, 65% and 25% contain sufficient NC to nations promoted an expansion of total cells (43 to 356-fold) and CFC (49 to 462-fold) within 7 days, however, engraft patients of 10 kg, 35 kg and 50-70 kg, respectively, when a threshold of 2 × 10 7 NC/kg body weight required early progenitors as defined by mixed-colony formation (CFU-GEMM) were substantially amplified only with for transplantation is assumed. Therefore, ex vivo expansion of stem/progenitor cells from human cord blood prior to SCF, Flt3-L plus IL-3 (94.3 ± 62.4-fold). H5100 medium or a serum-free medium supplemented with SCF, Flt3-L transplantation would be an attractive approach to obtain the quantity and quality of CB cells required for successful plus IL-3 were superior to 20% FCS/RPMI-1640 medium in the expansion of all progenitor cell types and were simi-(ie rapid, complete and sustained) long-term hematopoietic reconstitution after transplantation in adults. larly effective in supporting the amplification of total cells, CFC, CFU-GM, BFU-E/CFU-E and LTC-IC (maximumAlthough ex vivo expansion of CB cells has already been described, [10][11][12][13][14][15][16][17][18][19][20][21][22] in few reports all progenitor cell compartat day 7: 6.7 ± 3.4-fold and 5.5 ± 0.5-fold, respectively). However, the serum-free medium promoted a signifiments (ie LTC-IC) have been studied, to determine whether expanded CB cells still contain cells with long-term repopcantly higher expansion of CFU-GEMM (176.9 ± 81.7-fold) than H5100 medium (83.5 ± 26.2-fold) at day 7 and ulating potential. 10,18,19 This point is, however, of critical relevance for clinical application. Recently, a long-term only under serum-free conditions, CFU-GEMM were maintained over 14 days in tissue culture. These results expansion system was described, which allows a substantial amplification of primitive hematopoietic progenitors demonstrate that committed progenitors as well as the more immature CFU-GEMM and LTC-IC can be subincluding LTC-IC after 20 weeks in serum-containing liquid culture in the presence of Flt3-L and thrombopoiestantially amplified at the same time without exhausting the proliferative potential.tin. 19 However, for a clinically relevant expansion, a shorter culture period would be preferable. Thus, the establishment Keywords: cord blood; ex vivo expansion; Flt3-ligand of a short-term large-scale system for the simultaneous expansion of the various stem/progenitor cell populations involved in the different phases of ...

The development of a spreadable olive paste was carried out. Sevillian or Gordal green olive varieties were used in the paste. Pastes were prepared by crushing olives after pit removal and adding preservatives (potassium solvate and sodium benzoate), and antioxidants (T. B. H. Q.) to crushed olives. These were characterized by recording microbial, physical, chemical and sensory measurements and shelf life was evaluated over three months of storage at room temperature (18 °C) and under refrigeration (4 °C). The final microbial count of the paste was less than 100 ufc/ml. Sensory testing demonstrated that it was acceptable and storage conditions did not significantly affect the product.The pH of the paste decreased with time (4.3 to 4.17), resulting in increased acidity which was affected by storage temperature. The peroxide index, increased with time, (from 5 to 27 meq peroxide/kg oil). However, the pastes were not perceived to be rancid after sensory testing. Regarding the color, the main change observed was a darkening of the paste, as determined using the parameter L*. The shelf life of the olive paste was 90 days either under refrigeration or at room temperature.

Este estudio consistió en desarrollar una pasta untable de aceituna, variedad verde Sevillana o Gordal. La elaboración de la pasta consistió en deshuesar las aceitunas, triturarlas y adicionarles conservantes (sorbato de potasio y benzoato de sodio) y antioxidante (T. B. H. Q.), finalmente se envasaron. Ésta fue caracterizada microbiológica, físico-química y sensorialmente, y se evaluó su comportamiento durante 3 meses de almacenamiento a temperatura ambiente (18 °C) y en refrigeración (4 °C). El recuento microbiano de la pasta es menor a 100 ufc/ml. Sensorialmente el producto fue aceptado y esta condición no varió durante el almacenamiento, al igual que el contenido de aceite (70%) y a_w (0,90). El pH diminuye con el tiempo, por lo tanto, la acidez aumenta; viéndose afectado por la temperatura de almacenamiento. El índice de peróxido, aumenta en el tiempo (5 a 27 meq peróxido/kg aceite) y es mayor en condiciones ambientales, sin embargo, no es percibido como variación en la rancidez sensorial. En cuanto al color, el principal cambio producido es un oscurecimiento de la pasta expresado en el parámetro L*. La vida útil de la pasta de aceituna alcanza a 90 días, siendo apta su conservación a temperatura ambiente y de refrigeración