Adipose tissue stem cells (ADSCs) would be an attractive autologous cell source. However, ADSCs require invasive procedures, and has potential complications. Recently, urine stem cells (USCs) have been proposed as an alternative stem cell source. In this study, we compared USCs and ADSCs collected from the same patients on stem cell characteristics and capacity to differentiate into various cell lineages to provide a useful guideline for selecting the appropriate type of cell source for use in clinical application. The urine samples were collected via urethral catheterization, and adipose tissue was obtained from subcutaneous fat tissue during elective laparoscopic kidney surgery from the same patient (n = 10). Both cells were plated for primary culture. Cell proliferation, colony formation, cell surface markers, immune modulation, chromosome stability and multi-lineage differentiation were analyzed for each USCs and ADSCs at cell passage 3, 5, and 7. USCs showed high cell proliferation rate, enhanced colony forming ability, strong positive for stem cell markers expression, high efficiency for inhibition of immune cell activation compared to ADSCs at cell passage 3, 5, and 7. In chromosome stability analysis, both cells showed normal karyotype through all passages. In analysis of multi-lineage capability, USCs showed higher myogenic, neurogenic, and endogenic differentiation rate, and lower osteogenic, adipogenic, and chondrogenic differentiation rate compared to ADSCs. Therefore, we expect that USC can be an alternative autologous stem cell source for muscle, neuron and endothelial tissue reconstruction instead of ADSCs.
Adult Stem Cells/*cytology/*immunology/transplantation ; Biomarkers/metabolism ; Cell Differentiation ; Cell Lineage ; Cell Proliferation ; Cell Separation ; Chromosomal Instability ; Colony-Forming Units Assay ; Humans ; Karyotyping ; Multipotent Stem Cells/cytology/immunology/transplantation ; Subcutaneous Fat, Abdominal/*cytology ; Transplantation, Autologous ; Urine/*cytology

Chimerism testing permits early prediction and documentation of successful engraftment, and also facilitates detection of impending graft rejection. In this study, we serially monitored chimerism status by short tandem repeat-based PCR in nucleated cells (NC), T cells and natural killer (NK) cells after myeloablative allogeneic stem cell transplantation (SCT). Four patients with myeloid malignancies showed discrepant chimerism results among those three fractions. Three patients had mixed chimerism (MC) of donor/host T cells at a time point around the onset of chronic graft-versus-host disease (GVHD). In two patients with disease relapse, MC of NK cells preceded a morphological relapse or NK cells showed a higher percentage of patient cells compared to NC. Therefore, our study shows that chimerism analysis in lineage-specific cells might be useful in predicting clinical outcome after allogeneic SCT in certain patients.
Adult ; *Chimerism ; Graft vs Host Disease/*diagnosis/etiology ; Humans ; Killer Cells, Natural/cytology/immunology ; Male ; Microsatellite Repeats/*genetics ; Middle Aged ; Polymerase Chain Reaction ; Predictive Value of Tests ; Stem Cell Transplantation ; T-Lymphocytes/cytology/immunology ; Transplantation, Homologous

To investigate the characteristics and significance of reconstitution of peripheral blood plasmacytoid dendritic cell (PDC) precursors after allogeneic human leukocyte antigen mismatched/haploidentical hematopoietic stem cell transplantation, and its relationship with chronic graft versus host disease (cGVHD) and relapse, 19 patients with leukemia were enrolled for this study. Peripheral blood dendritic cell (DC) subsets of patients and healthy controls were detected by flow cytometry, and the correlations between reconstitution of DC and cGVHD, relapse were analyzed. The results showed that compared with healthy subjects, patients with leukemia had a significantly decreased proportion and absolute number of myeloid dendritic cell (MDC), MDC1, DC and the ratio of MDC/PDC (P<0.05). There were not statistically different in MDC2 and PDC between patients and healthy subjects. After transplant, all the proportion of WBC and absolute numbers of DC reached to healthy controls levels at 9 months (P>0.05), besides the proportion of PDC which reached to healthy controls levels at 1 year (P=0.494). Compared with levels before relapse, the proportions of MDC1, MDC, DC and the ratio of MDC/PDC were lower, but proportions of MDC2 and PDC were slightly higher after relapse. Patients with a 'high' PDC recovery profile had an improved cumulative incidence of cGVHD in contrast to patients with a 'low' PDC recovery profile on day 120 after transplantation (P=0.007). It is concluded that compared with healthy subjects, de novo leukemia patients have a significantly decreased proportion and absolute number of DC and the ratio of MDC/PDC before haploidentical hematopoietic stem cell transplantation; while ratio of MDC/PDC can be normalized with relative rapidity, the proportions of all DC subsets reached to normal levels on the whole at 9 months after transplantation, and also recovery level of DCs is correlated with occurrence of cGVHD and relapse.
Adolescent ; Adult ; Burkitt Lymphoma ; therapy ; Child ; Dendritic Cells ; cytology ; immunology ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; HLA Antigens ; immunology ; Haplotypes ; immunology ; Histocompatibility ; Histocompatibility Testing ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; therapy ; Male ; Middle Aged ; Myeloid Cells ; cytology ; immunology ; Peripheral Blood Stem Cell Transplantation ; adverse effects ; methods ; Plasma Cells ; cytology ; immunology ; Recurrence

To compare the expression of CD antigens on immune cells from umbilical cord blood (UCB) and bone marrow (BM) and analyze its clinical significance, the phenotypes of lymphoid cells and nucleated cells from 38 UCB and 10 BM samples were investigated by flow cytometry with double labeling monoclonal antibodies. The results showed that the immature lymphocytes (CD3(-) CD4(+)) were detected in UCB and higher than those in BM; cytotoxic T lymphocytes (CD3(+) CD16(+) CD56(+)) in UCB were significantly lower than those in BM. NK cells (CD3(-) CD16(+) CD56(+)) in UCB were higher than those in BM. The ratio of CD34(+) cells in nucleated cells of UCB was similar to that of BM, however, both the contents of myeloid (CD34(+) CD13(+) and CD34(+) HLA-DR(+)) and lymphoid (CD34(+) CD19(+)) progenitor cells in UCB were lower than those in BM. It is concluded that the immune cells in UCB possess immaturity, which might lead to mild GVHD after UCB transplantation. It is inferred from the higher ratio of NK cells in UCB, GVL will not decrease after UCB transplantation. The lower contents of myeloid and lymphoid progenitor cells in UCB probably accounted for the slow hematopoiesis and immune reconstitution following UCB transplantation.
Adult ; Antigens, CD34 ; analysis ; Bone Marrow Cells ; immunology ; physiology ; Female ; Fetal Blood ; cytology ; immunology ; Flow Cytometry ; Graft vs Host Disease ; etiology ; Hematopoiesis ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunophenotyping ; Infant, Newborn ; Lymphocytes ; immunology

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.
Adult ; Birth Weight ; Blood Banks ; methods ; Cord Blood Stem Cell Transplantation ; methods ; Delivery, Obstetric ; Donor Selection ; Female ; Fetal Blood ; cytology ; immunology ; Gestational Age ; Hematopoietic Stem Cells ; Humans ; Infant, Newborn ; Male ; Maternal Age ; Pregnancy ; Young Adult

This study was purposed to investigate the immune reconstitution of T-cells in patients who received haploidentical hematopoietic stem cell transplantation (hiHSCT). The peripheral blood was harvested from 22 patients before transplantation and at month 1, 3, 6 after hiHSCT. The proportions of T lymphocyte subtypes including CD3(+), CD4(+), CD8(+), CD45RO(+), and CD45RA(+)CD62L(+) were analyzed by flow cytometry, followed by the calculation of T cell numbers according to the amounts of peripheral blood leukocytes. Adenosine triphosphate (ATP) value in CD4(+) T cells was measured by ImmuKnow method to evaluate the function of lymphocytes. The results showed that the CD3(+) cell absolute value before transplantation was 833.75 ± 359.84/µl, but those values at month 1, 3, 6 after transplantation were 318.87 ± 266.71/µl, 1006.76 ± 512.32/µl and 1296.38 ± 958.77/µl respectively. The CD4(+) cell absolute value before transplantation was 336.99 ± 211.11/µl, but such values at month 1, 3, 6 after transplantation were 45.89 ± 44.21/µl, 142.97 ± 114.85/µl, and 181.78 ± 120.61/µl respectively. The CD8(+) cell absolute value before transplantation was 430.21 ± 159.48/µl, but those values at month 1, 3, 6 after transplantation were 230.44 ± 195.89/µl, 621.64 ± 318.83/µl, and 823.07 ± 633.55/µl respectively. The CD4(+)CD45RO(+) memory T cell absolute value before transplantation was 227.44 ± 73.34/µl, but such values at month 1, 3, 6 after transplantation were 43.47 ± 43.40/µl, 138.69 ± 110.17/µl, 147.73 ± 82.94/µl respectively. The CD8(+)CD45RO(+) memory T cell absolute value before transplantation was 212.70 ± 98.48/µl, but such values at month 1, 3, 6 after transplantation were 184.76 ± 168.65/µl, 445.90 ± 252.50/µl, 519.80 ± 475.53/µl respectively. CD4(+)CD45RA(+)CD62L(+) naive T cell number before transplantation was 68.94 ± 59.74/µl, but such cell numbers at month 1, 3, 6 after transplantation decreased to 2.44 ± 2.93/µl, 3.14 ± 3.48/µl, 23.22 ± 38.38/µl respectively. The CD8(+)CD45RA(+)CD62L(+) naive T cell absolute value before transplantation was 124.82 ± 60.95/µl, but those values at month 1, 3, 6 decreased to 19.37 ± 17.71/µl, 76.63 ± 50.85/µl, and 114.49 ± 174.29/µl respectively. The ATP value in CD4(+) T cells decreased to 210.19 ± 119.37 ng/ml at month 1 after transplantation and increased to 280.62 ± 110.03 ng/ml at month 3, and 357.28 ± 76.18 ng/ml at month 6 after transplantation. It is concluded that CD8(+) memory T cell reconstruction contributes critically to T cell recovery early after hiHSCT, while the thymic output function remains low. However, T cell function recovers to normal range at month 3 after transplantation.
Adolescent ; Adult ; CD8-Positive T-Lymphocytes ; cytology ; Child ; Child, Preschool ; Female ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunophenotyping ; Killer Cells, Natural ; immunology ; Lymphocyte Count ; Male ; T-Lymphocyte Subsets ; immunology ; Young Adult

The aim of this study was to investigate the biological characteristics of human bone marrow mesenchymal stem cells from bone marrow of different age donors. The experiments were divided into four groups by donors age, group A represented MSC derived from fetal bone marrow, group B represented MSC derived from bone marrow of 0-20 years old donors, group C represented MSC derived from bone marrow of 20-40 years old donors and group D represented MSC derived from bone marrow of donors older than 40. The growth, purification, proliferation and multipotential abilities of MSC in 4 groups were observed and their immunophenotypes were determined by flow-cytometry. The level of cytokines (IL-6, SCF, FLT-3L, SDF-1 and TGF-beta1) were assayed by ELISA method. Cell cycles were analyzed to show the proliferation index (PrI). MSCs derived from bone marrow of 4 groups were injected subcutaneous into NOD/SCID mouse to observe the safety. The results showed that different age donors bone marrow all gave rise to MSC. These cells were similar in morphology, antigenic phenotype, differentiation potential and cell cycle. The primary culture time of group B was shorter than other groups. The duration of passage 1 (P1) was 5.5 days, and the duration of P10 was 33 days, after P10 culture, (5.19 +/- 2.15) x 10(10) MSCs were obtained from 8 x 10(6) MNC of this group. The primary culture time of groups A, C, D were longer, the duration of P1 were 15, 7 and 13 days for group A, C and D respectively, and the duration of P10 was 50, 60 and 72 days for group A, C and D, respectively. After P10 culture, (4.98 +/- 2.08) x 10(10), (1.86 +/- 0.47) x 10(10), (0.64 +/- 0.22) x 10(10) MSCs were obtained from 8 x 10(6) MNC of group A, C and D respectively. The morphology of MSC of group A was longer and slender. The ability of expansion decreased after P15 for A group, P10 for B group and P8 for C and D groups. The levels of SCF, FLT3-L, IL-6 and SDF-1 in group B were higher than other groups. Karyotype analysis showed that MSCs from 4 groups were normal, and tumor-like tissues were not developed after cultured MSCs were inoculated in NOD/SCID mice. It is concluded that there was relationship between age and the biological characteristics of human bone marrow mesenchymal stem cells. For clinical use, especially in hematopoietic stem cell transplantation (HSCT), 0-20 years old donors were perfect MSCs donors who can provide sufficient MSCs in relatively short times. MSCs of group B can be used as stem cell source because the biological characteristics of MSCs of groups B are superior to that of other groups.
Adolescent ; Adult ; Age Factors ; Animals ; Blood Donors ; Bone Marrow Cells ; cytology ; immunology ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Child ; Child, Preschool ; Cytokines ; analysis ; Enzyme-Linked Immunosorbent Assay ; Fetus ; Flow Cytometry ; Humans ; Immunophenotyping ; Infant ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; immunology ; metabolism ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Transplantation, Heterologous

This study was aimed to explore the effect of donor characteristics (age, sex and so on.) on CD34(+) cell yields in apheresis from healthy donors mobilized by recombinant granulocyte colony-stimulating factor(rhG-CSF). In 61 healthy donors, the characteristics associated with CD34(+) cell yield were analysed. The relationship between the CD34(+) cell yields and donor characteristics was statistically assessed with multivariate forward, backward and stepwise regression methods. A variety of parameters were analyzed which included donor age, sex, weight, height, body mass index (BMI) and time for collection of peripheral blood apheresis, while the mean number of peripheral blood mononuclear cells (MNCs), CD34(+) cell count, CD34(+) cell proportion based on MNC and CD34(+) cell count per kg of donor weight were used as the variables. The results showed that age of donors was the main factor impacting CD34(+) cell yields (-0.60 < r < -0.45, p < 0.005). In a partial correlation analysis the body weight, height and BMI were served as control factors, the negative correlation of age with CD34(+) cell yields was still found (-0.50 < r < -0.35, p < 0.02). BMI was only weakly correlated with the yields of CD34(+) cells per kg(r = -0.297, p < 0.05). As a whole, sex showed no relation with the CD34(+) cell yields. Compared with the female group less than 35 years old, height, weight and BMI in male group of low age exerted a positive impact on CD34(+) cell yield. The optimal time for collection of PB was day 4 after treatment with rhG-CSF, when 70% of the donors could reach the peak CD34(+) cell yields. It is concluded that the age of the donors is the first factor determining the choice of donors for allogeneic hematopoietic stem cell transplantation, the sex, height, weight and BMI are secondary factors impacting yield of CD34(+) cells from donors mobilized with rhG-CSF.
Adolescent ; Adult ; Age Factors ; Antigens, CD34 ; immunology ; metabolism ; Blood Cell Count ; Blood Cells ; cytology ; immunology ; Body Height ; Body Mass Index ; Body Weight ; Child ; Female ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Recombinant Proteins ; Sex Factors ; Tissue Donors ; Young Adult

OBJECTIVETo apply autologous transplantation of peripheral blood stem cells in the treatment of critical limb ischemia.

METHODSFifteen patients with critical lower limb ischemia were recruited in this study. After bone marrow mobilization, peripheral blood stem cells were collected using CS-3000 Plus device, and transplanted directly into the ischemic limb. In total 17 times of transplantation were performed.

RESULTSOne year after implanted peripheral blood stem cells, the pain scale decreased from 5 to 0, the ankle-brachial pressure index (ABI) increased from 0.30 to 0.46, the pain-free walking distance and maximal walking distance increased from 0.15 to 0.72 km and 0.96 to 2.13 km separately, from a total of 6 of 15 limb ulcers of transplanted patients 5 were healed after cell transplantation.

CONCLUSIONAutologous peripheral blood stem cell transplantation is able to induce functional angiogenesis in ischemic limb, which can significantly reduce rest pain, increase walk distance, and promote ulcer healing.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; blood ; Arterial Occlusive Diseases ; surgery ; Arteriosclerosis Obliterans ; surgery ; Female ; Flow Cytometry ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; Hematopoietic Stem Cells ; cytology ; drug effects ; immunology ; Humans ; Leg ; blood supply ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; methods ; Treatment Outcome

Acute graft-versus-host disease (aGVHD) is a serious complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, the mechanisms of aGVHD are not well understood. We aim to investigate the roles of the three angiogenic factors: angiopoietin-1 (Ang-1), Ang-2 and vascular endothelial growth factor (VEGF) in the development of aGVHD. Twenty-one patients who underwent allo-HSCT were included in our study. The dynamic changes of Ang-1, Ang-2 and VEGF were monitored in patients before and after allo-HSCT. In vitro, endothelial cells (ECs) were treated with TNF-β in the presence or absence of Ang-1, and then the Ang-2 level in the cell culture medium and the tubule formation by ECs were evaluated. After allo-HSCT, Ang-1, Ang-2 and VEGF all exhibited significant variation, suggesting these factors might be involved in the endothelial damage in transplantation. Patients with aGVHD had lower Ang-1 level at day 7 but higher Ang-2 level at day 21 than those without aGVHD, implying that Ang-1 may play a protective role in early phase yet Ang-2 is a promotion factor to aGVHD. In vitro, TNF-β promoted the release of Ang-2 by ECs and impaired tubule formation of ECs, which were both weakened by Ang-1, suggesting that Ang-1 may play a protective role in aGVHD by influencing the secretion of Ang-2, consistent with our in vivo tests. It is concluded that monitoring changes of these factors following allo-HSCT might help to identify patients at a high risk for aGVHD.
Acute Disease ; Adolescent ; Adult ; Angiogenesis Inducing Agents ; immunology ; metabolism ; pharmacology ; Angiopoietin-1 ; genetics ; immunology ; pharmacology ; Angiopoietin-2 ; genetics ; immunology ; pharmacology ; Antineoplastic Agents ; therapeutic use ; Female ; Gene Expression Regulation, Neoplastic ; Graft vs Host Disease ; genetics ; immunology ; pathology ; Hematopoietic Stem Cell Transplantation ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; immunology ; Humans ; Leukemia, Myeloid ; genetics ; immunology ; pathology ; therapy ; Lymphoma, Non-Hodgkin ; genetics ; immunology ; pathology ; therapy ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; immunology ; pathology ; therapy ; Retrospective Studies ; Signal Transduction ; Transplantation, Homologous ; Tumor Necrosis Factor-alpha ; pharmacology ; Vascular Endothelial Growth Factor A ; genetics ; immunology