Molecular cytogenetic analysis of Korean patients with Waldenström macroglobulinemia.

To compare the molecular cytogenetic characteristics between Waldenström macroglobulinemia (WM) and multiple myeloma (MM), we performed interphase fluorescent in situ hybridization (FISH) in Korean patients with WM and MM. Forty patients with WM and 132 patients with MM were enrolled onto the study. FISH was performed with seven different probes: 6q21, 6q23, CEP4, CEP9, immunoglobulin (IgH) breakapart, RB1 gene, and 1q25. Out of 22 WM patients, 4 (18%) had abnormal karyotypes, mainly structural changes on conventional karyotyping. After performing FISH for the available 29 cases, deletions of 6q23 and 6q21 were newly detected in 3 cases (10%). There was no other anomaly, including trisomy 4 in WM. No 6q deletion was observed in MM patients, but RB1 deletion was the most common change (45%), followed by IgH translocation (42%) and gain of 1q (38%). In conclusion, Korean WM patients had a low rate of 6q deletion (10%) and no trisomy 4.

Hepatic differentiation of cord blood-derived multipotent progenitor cells (MPCs) in vitro.

Umbilical cord blood (UCB) is a rich source of hematopoietic stem cells that possesses practical and ethical advantages. We previously reported a novel UCB-derived adult stem cells which we termed umbilical cord blood-derived multipotent progenitor cells' (MPCs). MPCs were capable of differentiating into functional neuronal cells. Under appropriate conditions lasting several days or weeks, we now show that the MPCs differentiate into hepatocyte-like cells in vitro; their properties were verified using reverse transcription-polymerase chain reaction (RT-PCR), Western blot, immunofluorescence, periodic acid-Schiff (PAS) staining of accumulated glycogen and an enzyme-linked immunosorbent assay (ELISA). We also found that hepatic differentiated cells expressed hepatocyte specific markers, such as albumin, hepatocyte nuclear factor (HNF)-1alpha, HNF4, cytokeratin (CK)-8, CK-18, tyrosine amino transferase (TAT), and CYP2B6. Moreover, albumin was secreted, which suggests that MPCs from UCB possess multi-differentiation potential and have the capacity to differentiate into functional cells of hepatic lineage in vitro.

Neural differentiation of novel multipotent progenitor cells from cryopreserved human umbilical cord blood.

Umbilical cord blood (UCB) is a rich source of hematopoietic stem cells, with practical and ethical advantages. To date, the presence of other stem cells in UCB remains to be established. We investigated whether other stem cells are present in cryopreserved UCB. Seeded mononuclear cells formed adherent colonized cells in optimized culture conditions. Over a 4- to 6-week culture period, colonized cells gradually developed into adherent mono-layer cells, which exhibited homogeneous fibroblast-like morphology and immunophenotypes, and were highly proliferative. Isolated cells were designated 'multipotent progenitor cells (MPCs)'. Under appropriate conditions for 2 weeks, MPCs differentiated into neural tissue-specific cell types, including neuron, astrocyte, and oligodendrocyte. Differentiated cells presented their respective markers, specifically, NF-L and NSE for neurons, GFAP for astrocytes, and myelin/oligodendrocyte for oligodendrocytes. In this study, we successfully isolated MPCs from cryopreserved UCB, which differentiated into the neural tissue-specific cell types. These findings suggest that cryopreserved human UCB is a useful alternative source of neural progenitor cells, such as MPCs, for experimental and therapeutic applications.

Endothelial progenitor cell differentiation using cryopreserved, umbilical cord blood-derived mononuclear cells.

AIM: To investigate the endothelial differentiation potentiality of umbilical cord blood (UCB), we induced the differentiation of endothelial progenitor cells (EPC) from cryopreserved UCB-derived mononuclear cells (MNC). METHODS: MNC from cryopreserved UCB and peripheral blood (PB) were cultured in M199 medium with endothelial cell growth supplements for 14 d. EPC were characterized by RT-PCR, flow cytometry, and immunocytochemistry analysis. The proliferation of differentiated EPC was studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, and vascular endothelial growth factor (VEGF) concentration was measured using an ELISA kit. Characteristics of UCB-derived EPC were compared with those of PB-derived EPC. RESULTS: A number of round-shaped cells were loosely attached to the bottom after 24 h culture, and numerous spindleshaped cells began to appear from the round-shaped ones on d 7. Those cells expressed endothelial markers such as, Flt-1/VEGFR-1, ecNOS, VE-cadherin, von Willebrand factor, and secreted VEGF. The patterns of endothelial markers of EPC from PB and UCB did not show striking differences. The results of the proliferation and secretion of VEGF were also similar. CONCLUSION: We successfully cultured UCB cells stored at -196 Celsius degree into cells with the quality of endothelial cells. Those EPC could be used for angiogenic therapeutics by activating adjacent endothelial cells and enhancing angiogenesis.

Isolation of mesenchymal stem cells from cryopreserved human umbilical cord blood.

Umbilical cord blood (UCB) is well known to be a rich source of hematopoietic stem cells with practical and ethical advantages. Because mesenchymal stem cells (MSCs) from bone marrow have been regarded as good materials for cell/gene therapy as well as for tissue engineering because of their multidifferentiation potential, a number of trials have been undertaken to isolate MSCs from UCB. However, the results have been controversial, and little has become known about the effect of cryopreservation on the isolation of these stem cells. In this study, we examined the ability of cryopreserved UCB-derived cells to produce MSCs. Various culture conditions, including the seeding concentrations of cells and the media used, were investigated. We were able to obtain adherent cell populations after 3 to 5 weeks in our culture conditions from UCB-derived mononuclear cell fractions that had undergone cryopreservation for 0.1 to 5 years. These cells exhibited a fibroblast-like morphology and typical mesenchymal-like immunophenotypes. The results indicate that cryopreserved human UCB fractions can be used as an alternative source of MSCs for experimental and clinical applications as well as for tissue engineering.

Mesenchymal stem cells from cryopreserved human umbilical cord blood.

Umbilical cord blood (UCB) is well known to be a rich source of hematopoietic stem cells with practical and ethical advantages, but the presence of mesenchymal stem cells (MSCs) in UCB has been disputed and it remains to be validated. In this study, we examined the ability of cryopreserved UCB harvests to produce cells with characteristics of MSCs. We were able to obtain homogeneous plastic adherent cells from the mononuclear cell fractions of cryopreserved UCB using our culture conditions. These adherent cell populations exhibited fibroblast-like morphology and typical mesenchymal-like immunophenotypes (CD73+, CD105+, and CD166+, etc.). These cells presented the self-renewal capacity and the mesenchymal cell-lineage potential to form bone, fat, and cartilage. Moreover, they expressed mRNAs of multi-lineage genes including SDF-1, NeuroD, and VEGF-R1, suggesting that the obtained cells had the multi-differentiation capacity as bone marrow-derived MSCs. These results indicate that cryopreserved human UCB fractions can be used as an alternative source of MSCs for experimental and therapeutic applications.

High dose therapy followed by autologous peripheral blood stem cell transplantation as a first line treatment for multiple myeloma: a Korean Multicenter Study.

We conducted a phase II multicenter trial to estimate the response and survival of patients with newly diagnosed multiple myeloma to high dose melphalan therapy followed by autologous peripheral blood stem cell transplantation. Eligible patients who had undergone induction with vincristine, adriamycin and dexamethasone (VAD) should have adequate cardiac, pulmonary and renal function (creatinine <2 mg/dL). Melphalan at 200 mg/m2 was used as a conditioning regimen. Eighty patients were enrolled from 13 centers. The median age of the patients was 53 yr (range; 20 to 68 yr). The initial stage was IA/IIA/IIB/IIIA/IIIB in 3/8/1/54/14 patients, respectively. Beta2-microglobulin, CRP and LDH were increased in 74, 42 and 34% of the patients examined. Cytogenetic data were available in 30 patients, and 6 patients showed numeric or structural abnormalities. Two therapy-related mortalities occurred from infection. Among the 78 evaluable patients, CR/PR/MR/NC/PD were achieved in 48/26/2/1/1 patients, respectively. After a median follow-up of 30 months, the median overall and event-free survivals were 66 months (95% CI: 20-112) and 24 months (95% CI: 18-29), respectively. This study verifies the efficacy and feasibility of high dose melphalan therapy with autologous stem cell transplantation in newly diagnosed multiple myeloma.

Development and Test of an Information Needs Scale for Outpatients Undergoing Chemotherapy.

PURPOSE: The purpose of this study was to develop and test an Information Needs Scale for Korean outpatients undergoing chemotherapy (INS-C). MATERIALS AND METHODS: Thirty-three items of the INS-C had content validity based upon findings in the literature and the experiences of expert oncology physicians and nurses. Each item consisted of a five-point Likert scale from 1 (don't want to know) to 5 (want to know very much). The items were administered to 175 Korean outpatients undergoing chemotherapy. The data obtained was analysed using a factor analysis for construct validity and Cronabch's alpha for internal consistent reliability. RESULTS: From the factor analysis, six subscales were derived significantly. The six subscales explained 64.62% of the variance. The subscales were named Side-Effects/Investigative Tests (9 items), Spread of Disease (4 items), Financial Cost (2 items), Treatment (7 items), Activities/ Eating (6 items), and Interrelationships/Support (5 items). The Cronbach's alpha of the total INS-C was .95, and the alpha of the subscales ranged from .77 to .91. CONCLUSION: The present study suggests that the INS-C is a reliable and valid instrument to measure the information needs of outpatients undergoing chemotherapy. Health professionals caring for patients with cancer should assess the informational needs of their patients using a reliable and valid instrument and be prepared to provide accurate information.