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Umbilical cord blood (UCB) is an alternative source of hematopoietic stem cells for transplantation with success being associated with the total nucleated cell (TNC) count, CD34(+) cells and colony-forming unit-granulocyte-macrophage (CFU-GM) content infused. This study was purposed to clarify the impact of maternal and neonatal factors on hematopoietic potential of UCB product. UCB samples were screened, processed, tested and cryopreserved according to the Standard Operation Procedure (SOP) of Guangzhou cord blood bank (GZCBB). Relationship of hematopoietic cell parameters with maternal and neonatal characteristics for 4615 UCB units was analyzed retrospectively. The results showed that both collected volume (Mean ± SD: 95.23 ± 22.42 ml; Median: 91.85 ml) and initial TNC [Mean ± SD: (1.34 ± 0.49) × 10(9); Median: 1.25 × 10(9)] correlated well with postprocessed TNC [Mean ± SD: (1.21 ± 0.42) × 10(9); Median: 1.14 × 10(9); p < 0.001], CD34(+)count [Mean ± SD: (5.14 ± 4.55) × 10(6); Median: 4.08 × 10(6); p < 0.001] and CFU-GM content [Mean ± SD: (9.72 ± 8.66) × 10(5); Median: 7.53 × 10(5); p < 0.001]. As for donor factors, only infant birth weight correlated strongly with volume collected and all hematopoietic cell parameters (p < 0.001). UCB samples from bigger babies had higher collected volume, TNC, CD34(+) count and CFU-GM content (p < 0.001). Mother's age had no correlation with all the above parameters. Gestational age correlated positively with initial/postprocessed TNC (p < 0.001) and negatively with CD34(+) count (p = 0.04), but no relation with collected volume and CFU-GM content. Cesarean section produced superior volume (Mean ± SD: 97.05 ± 22.23 ml vs 92.53 ± 22.43 ml; Median: 94.08 ml vs 88.82 ml; p < 0.001), but inferior cell count than vaginal delivery (p < 0.001). Male infants had more initial volume and CD34(+) count (Mean ± SD: 96.41 ± 22.31 ml vs 93.95 ± 22.47 ml; Median: 93.27 ml vs 90.14 ml; p < 0.001); [Mean ± SD: (5.28 ± 5.04) × 10(6) vs (5.00 ± 3.94) × 10(6); Median: 4.18 × 10(6) vs 3.94 × 10(6); p < = 0.042], but lower initial and postprocessed TNC than female ones [Mean ± SD: (1.31 ± 0.50) × 10(9) vs (1.37 ± 0.47) × 10(9); Median: 1.22 × 10(9) vs 1.28 × 10(9); p < 0.001]; [Mean ± SD: (1.18 ± 0.42) × 10(9) vs (1.24 ± 0.41) × 10(9); Median: 1.10 × 10(9) vs 1.17 × 10(9); p < 0.001], while no significant difference of CFU-GM were found between male and female infants. It is concluded that these data may be helpful to optimize the UCB donor selection and improve cost efficiency of UCB bank resource. The heavier infants after vaginal delivery should be selected and large-volume units with higher TNC should be chosen at first.

The study was aimed to establish a protocol of isolating and culturing adult mesenchymal stem cells (MSC) from human bone marrow aspirate and identify them by surface antigen analysis and committed differentiation in order to provide an experimental foundation for achieving a therapeutic benefit in applying MSC in hematopoietic stem cell transplantation. MSCs were obtained from fresh human bone marrow aspirate by gradient centrifugation with Percoll (1.073 g/ml) and anchoring culture in L-DMEM with 10% fetal bovine serum by a full medium exchange every 3 days. The MSC surface antigens, including CD34, CD45, CD73, CD105, CD166, were analyzed on FACScan flow cytometer. Under culture in conditioned medium for osteogenesis (the hormone cocktail containing 0.1 micromol/L dexamethasone, 10 mmol/L glycerol-2-phosphate and 50 micromol/L ascorbic acid) and adipogenesis (the cocktail containing 1 micromol/L dexamethasone, 5 mg/L insulin, 0.5 mmol/L 1-methyl-3-isobutylxanthine and 60 micromol/L indomethacin), MSCs committedly differentiated into osteoblasts and adipocytes. The differentiated mesenchymal stem cells were identified by morphological observation and immunohistochemical staining. The results showed that by gradient centrifugation and adhesion culture, MSCs could be isolated and culture-expanded from human bone marrow aspirate. These cells were uniformly negative for CD34, CD45 and positive for CD73, CD105 and CD166. The osteogenic differentiated cells were positive for alkaline phosphatase (ALP) and the adipogenic differentiated cells displayed accumulation of lipid vacuoles, as detected by oil red O. It is concluded that MSC can be isolated and expand-cultured from adult human bone marrow aspirate and committedly differentiate into osteoblasts and adipocytes. MSC primary identification can be accomplished by flow cytometry and induced differentiation. The set of methods in current experiment shows somewhat practical value for basic research and clinical application.

To study the program of evaluating mothers and infants after 6 months of cord blood donation, from June 1998 to February 2004, all mothers after 6 months of cord blood donation were followed-up by phone calls or letters to report on the health condition. The results showed that when 3 195 mothers were visited by phone calls, 18 mothers declined to answer. 392 letter were send to those who could not be found by phone, 15 of whom wrote back. The average time to talk with each mother was approximately 12 minutes. Follow-up on the baby donors showed two cases with chromosome abnormality, one with hypothyroidism, one with neutropenia, one with albinism and 5 dead with unclear reasons. The cord blood components from all these abnormal donors found were discarded. In conclusion, the programs to evaluate mother and baby after 6 months of cord blood donation seems important in quality control of the components stored in cord blood bank.

In order to research the related factors of umbilical cord blood transplantation, 54 cases of unrelated umbilical cord blood transplantation were analyzed retrospectively, which were performed from June 1998 to July 2003. All cord blood units were obtained from full term normal vaginal deliveries in Guangzhou Maternal-Neonatal Hospital. The fractionation, cryopreservation and thawing of cord blood have been done according to the regulation of New York umbilical cord blood bank and pertinent literature. The selection of cord blood is based on HLA typing and the number of nucleated cells. The results showed that from June 1998 to July 2003, 3 475 units of cord blood were collected in Guangzhou Umbilical Cord Blood Bank and 99 units were provided for therapy of 85 patients in 21 transplantation centers, including 11 sibling and 74 unrelated cord blood transplantations. 54 cases of unrelated cord blood transplantation were reported, including 43 malignant diseases and 11 non-malignant diseases. The median age of recipients was 9.5 (range 1.2 - 33) years, the median weight was 27 (range 10 - 60) kg, the median number of TNC was 6.82 x 10(7)/kg, 43 cord blood were implanted (ANC > 500/microl) at day 60 after transplantation (79.6%, median 17). The time of nuclear cell reconstitution after cord blood transplantation was statistically related with nucleated cells and the type of disease, not related with HLA matching. Acute GVHD was present in 8 patients (21.6%) and chronic GVHD occurred in 2 patients (5.4%), 6 patients suffered from graft failure (11.1%). The total survival rate was 42.6%. It is suggested that unrelated umbilical cord blood transplantation seems to be a good substitute for bone marrow transplantation and has good prospects especially in children.

In order to study the feasibility of multi-umbilical cord blood transplantation (multi-UCBT) in adult, 13 cases of unrelated allogeneic multi-UCBT performed from June 1998 to July 2003 were analyzed retrospectively. All cord blood units were obtained from full term normal vaginal and cesarean deliveries in Guangzhou Maternity and Neonatal Hospital. The fractionation, cryopreservation and thawing of cord blood were done according to the regulation of New York Umbilical Cord Blood Bank and pertinent literatures. Donors of HLA 1/6-2/6 mismatch were accepted at registry search. The results showed that from June 1998 to July 2003, 28 umbilical cord blood units were selected by 7 transplantation centers for 13 cases. The median age of recipients was 22 (8 - 41) years, and the median weight was 50 (21 - 75) kg, the median infused dose of total nuclear cells was 2.91 x 10(7)/kg. Six out of thirteen cases were engrafted after cord blood infusion with absolute neutrophil count of > 5.0 x 10(8)/L at 19 days post-infusion. Only one case suffered from graft versus host disease, the total survival of multi-UCBT was 46.2% (6/13). It is concluded that good prospects in the field of multi-umbilical cord blood transplantation is likely to be realized.

The objective of this research was to explore whether the number of CD34(+)CD38(+) cells infused affects hematopoietic reconstitution after cord blood transplantation. The number of CD34(+)CD38(+) cells in cord blood was analysed with flow cytometry after freezethawing. The body weight and time for neutrophil and platelet recovery were measured in 20 children with acute leukemia. The results showed that the median number of CD34(+)CD38(+) cells infused was 29.47 (9.85 - 325.71) x 10(4)/kg. A median time for neutrophil recovery (> 5 x 10(8)/L) in 20 patients was 18.5 (11 - 32) days, and time for platlet recovery (> 2 x 10(10)/L) in 19 of 20 patients was 45 (12 - 118) days. The number of CD34(+)CD38(+) cells infused correlated with time to neutrophil and platelet recovery (r = -0.577, P < 0.01 and r = 0.503, P < 0.05, respectively). In conclusion, the number of CD34(+)CD38(+) cells infused is correlated with the time for hematologic recovery.