Source Of Stem Cells For Transplantation Research Articles

Twenty years of experience on stem cell transplantation in iran.

BackgroundHematopoietic stem cell transplantation (HSCT) is a new window to therapy of many diseases. From March 1991 through April 2011, a total of 3237 HSCT were performed in the Hematology-Oncology and Stem Cell Transplantation Research Center, affiliated to Tehran University of Medical Sciences. Here we report 20 years experience of HSCT.ObjectivesOur strategy and aim include the protraction of cytogenetic and molecular biological diagnostic tests, the expansion of the first Iranian Cord Blood Bank (ICBB) and development of the first Iranian Stem Cell Donor Program (ISCDP), and improvement the researches in new therapeutic fields.Patients and MethodsTotally, 3237 patients were undergone HSCT. Of these transplants, 2205 were allogeneic stem cell transplantation, 1016 autologous and 16 syngeneic. Among 2205 patients who were undergone allogenic-HSCT, 34 received cord blood stem cells as stem cell source for transplantation. It is important to point out that cord blood bank at our center provides reliable storage of cord blood stem cells for our patients. Stem cell transplantation was performed for treatment of various diseases such as acute myelogenous leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphoblastic leukemia, beta-thalassemia major, sickle- cell thalassemia, sickle- cell disease, multiple myeloma, myelodysplasia, mucopolysaccharidosis, paroxysmal nocturnal hemoglobinuria, non-Hodgkin’s lymphoma, Hodgkin’s disease, severe aplastic anemia, plasma cell leukemia, Niemann-Pick disease, Fanconi anemia, severe combined immunodeficiency, congenital neutropenia, leukocyte adhesion deficiencies, Chediak-Higashi syndrome, osteopetrosis, histiocytosis X, Hurler syndrome, amyloidosis, systemic sclerosis, breast cancer, Ewing's sarcoma, testicular cancer, germ cell tumors, neuroblastoma, medulloblastoma, renal cell carcinoma, nasopharyngeal carcinoma, ovarian cancer, Wilms’ tumor, rhabdomyosarcoma, pancreatoblastoma, and multiple sclerosis. Also, we had 220 cellular therapies for post-myocardial infarction, multiple sclerosis, cirrhosis, head of femur necrosis, Diabetes Mellitus and GvHD treatment. 45 patients were undergone retransplantation in this center.ResultsAbout 78.2% of the patients (2530 of 3237) remained alive between one to 211 months after stem cell transplantation. Nearly, 21.8% (707) of our patients died after stem cell transplantation. The main causes of death were relapse, infection, hemorrhagic cystitis, graft-versus- host disease and etc.ConclusionsIn Iran, HSCT has been successfully adapted in routine clinical care. Recently, new methods such as double cord blood and haploidentical transplantation have been used to treat many life-threatening diseases.

Rapid and Potent Mobilization of Murine Hematopoietic Stem and Progenitor Cells by the Novel CXCR4 Antagonist POL5551

AbstractAbstract 4100 INTRODUCTION:Mobilized hematopoietic stem/progenitor cells (HSPC) have become the favored cell source for stem cell transplantation. The current gold standard mobilizing agent is G-CSF, where a 5-day mobilization regimen precedes stem cell harvest. More fast-acting and potent mobilizing agents would be desirable in the interest of donor and recipient safety and convenience. AMD3100, a currently available fast-acting mobilizing agent has proven weak for clinical mobilization as a single agent, with an efficiency of less than 1/5th of G-CSF in humans. METHODS:Binding properties (position, selectivity, affinity) of the novel PEM CXCR4 antagonist POL5551 to its target receptor were analyzed. In vivo mobilization efficiency was studied after injection into C57Bl/6 or DBA/2 mice. Different administration modes (Bolus vs. continous infusion) were tested as well as a combination with a standard regimen (9×100 μg/kg q12h) of G-CSF or Cyclophosphamide. Progenitor cell mobilization was monitored using clonogenic in vitro assays. Properties of mobilized cells were tested by flow cytometry, in vitro transwell migration assays and in vivo (homing, engraftment kinetics, stem cell contents, secondary engraftment) in lethally irradiated CD45.1 or CD45.1/2 recipient mice, alone or with CD45.1 competitor bone marrow cells. RESULTS:POL5551 showed selective binding to CXCR4 with an affinity exceeding that of its natural ligand SDF1, albeit occupying the extracellular receptor domains only (Fig.1). Mobilization peaked 4 hours after i.p. injection and a positive but non-linear dose-response relationship was documented for doses between 0.5 and 100 mg/kg (6000 CFU-C/ml, Fig. 2). A dose of 15 mg/kg mobilized more than twice the number of CFU-C as an equimolar dose of AMD3100, and a single dose of POL5551 at 30 mg/kg mobilized as strongly as a standard 5-day course of G-CSF treatment. POL5551 synergized with G-CSF in that injection of 5 mg/kg POL5551 after G-CSF treatment increased mobilization by 10-fold (3,000 to approx. 30,000 CFU-C/mL); this represents a 2.5 fold increase compared to a similar treatment regimen with AMD3100. Similarly, synergism with Cyclophosphamide was observed (9,900 to 50,000 CFU-C/mL).Given as continous infusion, 5 mg/kg/day of POL5551 mobilized up to 8,000 CFU-C/ml, whereas at 30 mg/kg/day up to 40,000 CFU-C/ml were measured in circulation on day 3. Mobilized cells migrated efficiently in in vitro transwell assays and homed efficiently to the bone marrow of lethally irradiated recipients. Moreover POL5551 mobilized cells provided timely early engraftment and contained long-term engrafting stem cells with self-renewal capacity, including in serial transplantation. The immunophenotype of immature cells mobilized with POL5551 was characterized by low expression of several adhesion molecules. CONCLUSIONS:POL5551 mobilizes murine stem and progenitor cells with rapid kinetics and unprecedented efficiency, markedly exceeding that of G-CSF and AMD3100. The combination of POL5551 with G-CSF mobilized more strongly than G-CSF with other CXCR4 antagonists. Similar to what we previously described for other mobilized stem cell specimen, POL5551-mobilized cells homed to marrow and engrafted efficiently. Immunophenotype was similar to that of AMD3100 mobilized cells. If the data can be corroborated in humans, POL5551 has the potential to substitute for G-CSF as a mobilizing agent. [Display omitted] [Display omitted] Disclosures:Romagnoli:Polyphor Ltd.: Employment. Chevalier:Polyphor Ltd: Employment. Patel:Polyphor Ltd.: Employment.

Comparison of the effects of human adipose and bone marrow mesenchymal stem cells on T lymphocytes

Mesenchymal stem cells (MSCs) are multipotent cells that can be derived either from the bone marrow (bMSCs) or adipose tissue (aMSCs). We have compared the immune regulatory properties of cells derived from bone marrow and adipose tissue to provide a theoretical basis for the choice of stem cell source for transplantation. The phenotypes of bMSCs and aMSCs are similar, differing only in the expression of CD106. aMSCs proliferate faster than bMSCs, but aMSCs suppressed T-lymphocyte proliferation and activation more poorly than bMSCs. Thus cell origin and abundance are important factors in determining the suitability of MSCs for transplantation. Adipose tissue offers a more promising source of cells for such an application.

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Aim We have recently established a cord blood bank at KAIMRCaiming to use these units for stem cell transplantation at our center and worldwide. Our aim was to investigate the frequency of HLA alleles in cord blood units collected at KAIMRC and to compare them with the HLA frequency from normal healthy Saudis. Methods Blood was collected from cord blood units from consented mothers and DNA was extracted. HLA types were determined using the polymerase chain reaction sequence-specific oligonucleotide (PCR-SSP)method for previously typed healthy controls and by Luminex SSO technology for 163 cord blood units. All samples were typed for HLA-A, HLA-B and HLA-DRB1. Results The most frequent HLA alleles in both cord blood units and healthy control subjects were HLA-A ∗ 02, HLA-B ∗ 50, HLA-B ∗ 51, HLA-DRB1 ∗ 04 HLA-DRB1 ∗ 07 and HLA-DRB1 ∗ 13. High resolution HLA-DRB1 typing results revealed that DBB1 ∗ 07 was of limited polymorphism all individuals were HLA-DRB1 ∗ 07:01 while for DRB1 ∗ 04 and DRB1 ∗ 13 many alleles were detected including HLA-DRB1 ∗ 04:01⧹02⧹05⧹06⧹13⧹30⧹36⧹59 and HLA-DRB1 ∗ 13:01⧹02⧹03⧹22⧹34⧹39⧹69⧹79⧹96. HLA-DRB1 ∗ 04:05 and HLA-DRB1 ∗ 13:02 were the most common alleles. Conclusions Our collected cord blood units represent the Saudi population and is a useful source for stem cell transplantation.

Predictors of the Development of Surgical Complications among Hematopoietic Stem Cell Transplantation Recipients

This study was designed to determine possible risk factors for the development of surgical complications after hematopoietic stem cell transplantation (HSCT). HSCT carries the possibility for the development of surgical morbidities and mortalities; certain populations of patients are at higher risk for developing complications. Defining those risk factors will help surgeons to anticipate and manage these complications. The records of 165 patients at King Hussein Cancer Center (KHCC) were reviewed from January 2007 to December 2008. Patients' characteristics, including age, source and type of transplant, behavior of disease, total body irradiation (TBI), immunity status, and intensity of conditioning were studied. According to our data, abdominal complications were more common among patients who received TBI versus patients who did not, and in patients who received peripheral blood versus bone marrow and cord blood as the source of stem cells. Ear, nose, and throat (ENT) complications were found more in patients who underwent nonmyeloablation conditioning. Catheter-related complications were found more significantly in immune-compromised patients during the post-engraftment period. We also found that ENT complications were more frequent in patients with cord blood as the source of stem cells, although the results are inconclusive due to the small sample size. Pretransplantation factors can be used as predictors for the development of surgical complications in HSCT recipients. Abdominal complications were significantly more common in patients who received total body irradiation as part of their conditioning regimen and when the peripheral blood was a source for stem cell transplantation.

The Novel Use of Hyperbaric Oxygen Therapy to Improve Umbilical Cord Blood Stem Cell Engraftment in Irradiated NOD/SCID Mice: What Did We Learn?

Abstract 4695 Introduction:UCB is also an attractive stem cell source for transplantation because of the ease of procurement, the absence of donor risks, and reduced risk of transmissible infections. Unfortunately, outcomes of UCB stem cell transplantation are less than optimal because of low cell dose in umbilical cord blood units and defects in homing to bone marrow, which result in both delayed engraftment and delayed immune recovery, both of which increase the mortality rate in UCB transplants. In these experiments, we investigated the use of hyperbaric oxygen therapy (HBO) to improve cord blood stem cell homing and engraftment post irradiation. Method:Six to eight week old NOD/ SCID mice were sublethally irradiated 24 hours prior to cord blood stem cell infusion. The irradiated mice were then divided into two groups (10 mice each). One group of mice served as the control group were mice remained under normoxic conditions. The second group of animals, the HBO group, six hours prior to cord blood stem cell infusion, were exposed to two hours of hyperbaric conditions in a hyperbaric chamber, with 100% oxygen for 2 hours at 2.5 atmospheres absolute (ATA). Erythropoietin (epo) level was checked by ELISA, at time of cord blood stem cell infusion, and was used as a surrogate marker for effective HBO therapy. Prior to CD34+ cord blood stem cell injection, cryopreserved cord blood units obtained from St. Louis Cord Blood Bank were processed for CD34+ selection followed by their transduction with a luciferase lentivirus so that they can be serially imaged, using Xenogen-IVIS imaging system, for engraftment into the bone marrow. Subsequently, cord blood stem cells (7 × 104 / 100 uL) in sterile PBS were administered by tail vein injection. The mice were weighed every other day and were sacrificed at 9–10 weeks post cord blood stem cell infusion. Peripheral blood and spleen tissue were obtained to determine donor engraftment by flow cytometry. Bone tissue was harvested for immunohistochemistry staining for human CD45. Result:Mean epo level in the HBO group was significantly lower than the mean epo level in the control group (226.7 ± 18.02 versus 294.4 ± 14.35 pg/ml (P=.0148)), consistent with effective HBO therapy. The average photon emission, or radiance, measured using the average of ventral and dorsal photon emission values per group, starting week 5, peaked at 7 weeks in the control mice, while peaked at 5 weeks in the HBO group. Since only ventral photon emission values were available during the first 4 weeks, an earlier peak in HBO group could not be excluded. One mouse in each group died, which was thought to be secondary to excessive bleeding related to mandible bleeding. No significant differences in median percentage peripheral blood or spleen engraftment were detected between the two groups. Interestingly, the HBO mice demonstrated a higher mean body weight started day 36 post cord blood stem cell infusion and continued until the end of study. Conclusions: HBO treatment did not result in detectable differences in terms of mean percentage peripheral blood or spleen engraftment at the end of study. However, HBO treated mice, compared to controls, showed an earlier photon emission peak values followed by lower photon emission values overtime. This earlier peak might have corresponded to the significant increase in weight noticed in the HBO mice starting week 5 post cord blood stem cell infusion.*We acknowledge the help we received from St. Louis Cord Blood Bank who provided the cryopreserved cord blood units for this research. Disclosures:No relevant conflicts of interest to declare.

Feasibility and Safety of Adoptive Immunotherapy with CIK Cells after Cord Blood Transplantation

Five patients with aggressive acute leukemias who had relapsed after cord blood transplantation were treated with cord blood derived cytokine-induced killer (CIK) cells. These were obtained by ex vivo expansion, using as starting material the washouts of the cord blood units, left over at the end of the transplant. We did not observe any acute or delayed adverse event, and observed 1 partial response in 1 patient concomitantly with the development of acute grade III graft-versus-host disease (GVHD). These observations show the relatively low toxicity of cord blood-derived CIK cells and, more importantly, the feasibility of this immunotherapy program for patients who could not otherwise benefit from donor lymphocyte infusions.

Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution

Delayed myeloid engraftment after cord blood transplantation (CBT) is thought to result from inadequate numbers of progenitor cells in the graft and is associated with increased early transplant-related morbidity and mortality. New culture strategies that increase the number of cord blood progenitors capable of rapid myeloid engraftment after CBT would allow more widespread use of this stem cell source for transplantation. Here we report the development of a clinically relevant Notch-mediated ex vivo expansion system for human CD34(+) cord blood progenitors that results in a marked increase in the absolute number of stem/progenitor cells, including those capable of enhanced repopulation in the marrow of immunodeficient nonobese diabetic-severe combined immunodeficient (NOD-SCID) mice. Furthermore, when cord blood progenitors expanded ex vivo in the presence of Notch ligand were infused in a clinical setting after a myeloablative preparative regimen for stem cell transplantation, the time to neutrophil recovery was substantially shortened. To our knowledge, this is the first instance of rapid engraftment derived from ex vivo expanded stem/progenitor cells in humans.

Umbilical Cord Blood Transplantation for Acute Myeloid Leukemia

Allogeneic hematopoietic stem-cell transplantation (HSCT) from a matched related or unrelated donor can cure an adult with relapsed or high-risk acute myeloid leukemia (AML). However, because many of these patients lack a suitable donor, umbilical cord blood has been explored as a stem-cell source for transplantation. In a retrospective analysis of international transplantation registries, researchers identified …

Rapid Mobilization of Murine Hematopoietic Stem and Progenitor Cells with TG-0054, a Novel CXCR4 Antagonist.

Abstract 3542Poster Board III-479 BackgroundThe interaction between SDF-1 and its receptor, CXCR4, is responsible for retaining of stem cells in the bone marrow. CXCR4 antagonist disrupts the SDF-1/CXCR4 interaction and mobilizes CD34+ hematopoietic stem cells (CD34+ HSCs) and CD133+ endothelial progenitor cells (CD133+ EPCs) from bone marrow into circulation, which can be used as a source for stem cell transplantation and other potential clinical indications. TG-0054 is a novel CXCR4 antagonist. In vitro CXCR4 antagonistic activity and in vivo stem cell mobilization activity of TG-0054 were determined in this study. Materials and MethodsIn vitro pharmacological assays, including GTP-binding assay, calcium mobilization assay, and chemotaxis assays, were performed to assess the potency of TG-0054 as a CXCR4 antagonist. In addition, receptor-binding assays against a panel of human chemokine receptors as well as other 68 cellular receptors were screened to evaluate the specificity of TG-0054. Kinetics and dose-dependent response of stem cell mobilization by TG-0054 were demonstrated in BALB/c mice. Activity of stem cell mobilization of TG-0054 when used alone or combined with G-CSF was also studied. ResultsTG-0054 blocked SDF-1-binding to CXCR4 receptor with an IC50 of 10 nM and inhibited SDF-1–induced GTP-binding (IC50 = 6 nM), chemotaxis (IC50 = 43 nM), and calcium mobilization (IC50 = 59 nM). TG-0054 showed greater than 3000-fold selectivity for CXCR4 receptor over other chemokine receptors. It is noteworthy that the IC50 for the inhibitory effect of TG-0054 on hERG potassium currents was greater than 1000 μM when human embryonic kidney (HEK) cells were used to examine the in vitro effects on the hERG potassium channel currents. In animal studies, TG-0054 rapidly and effectively mobilized CD34+ HSCs and CD133+ EPCs into circulation. Single intravenous (IV) administration of TG-0054 resulted in a rapid increase of total white blood cell (WBC) counts, as well as CXCR4+, CD34+, and CD133+ cells in peripheral blood between 1–3 hour post-injection and the cell counts returned to baseline within 6 hours. At their maximum tolerated doses (MTD), TG-0054 increased CXCR4+ cells by 28.7-fold in mice. Furthermore, TG-0054 efficiently mobilized CD34+ (14.5-fold) and CD133+ (7.9-fold) cells. The combined effects of TG-0054 and G-CSF on HSCs and EPCs mobilization from the bone marrow in mice were also investigated. G-CSF (100 μg/kg/day) was administered subcutaneously (SC) from Day 1 to Day 4 followed by a single IV injection of TG-0054 or AMD3100 on Day 5. Synergistic effects were observed in all cell types in mice receiving combination of G-CSF and 50 mg/kg of TG-0054 (total WBC 23.0-fold, CXCR4+ 29.0-fold, CD34+ 37.1-fold, CD133+ 110.8-fold of increase in combination group). ConclusionIt was concluded that TG-0054 was a potent and selective CXCR4 antagonist intended for the use of stem cell transplant and other clinical indications. It showed strong stem cell mobilization activity comparable to G-CSF when used alone, and demonstrated synergistic effects when combined with G-CSF in a nonclinical model. Disclosures:Huang:TaiGen Biotechnology Inc.: Employment. Liu:TaiGen Biotechnology Inc.: Employment. Yen:TaiGen Biotechnology Inc.: Employment. Chen:TaiGen Biotechnology Inc.: Consultancy. Chen:TaiGen Biotechnology Inc.: Consultancy. King:TaiGen Biotechnology Inc.: Employment. Hsu:TaiGen Biotechnology, Inc.: Membership on an entity's Board of Directors or advisory committees.

Survey On Directed Family Cord Blood Banking for Transplantation Among the National Cord Blood Bank Network in Italy.

Abstract 4195Directed family cord blood (DCB) storage provides hemopoietic stem cell source for transplantation (HSCT) for families with an existing or a potentially future recipient with HSCT curable disease (D). The National CB Banks Network in Italy (ITCBN) has a leading role in providing public DCB service for high-risk families, in compliance to GITMO directives for eligibility criteria (HSCT curable D: malignant MD, non MD, inherited ID). To provide best cost-effective practices recommendations it is important to report on DCB procedures and HSCT rate (HSCT-R) among public Banks. By 12.12.2008 almost 1800 DCB units were stored in 18 Italian Banks and 104 (9%) issued for HSCT. The present survey aims at summarizing the over 15 yrs DCB experience among 5 ITCBN Banks active since 1997 (range 1990-1997), and including 670 DCB units. ResultsPreliminary analysis reports a 94% overall compliance to eligibility criteria directives, and overall HSCT- R for an alive sibling of 12% (63/522); the 63 HSCT were 97% matched, for curing ID in 84% and with 72 % overall survival outcome. Different policies among Banks were compared (Bank vs others: 1) eligibility criteria distribution : Bank PV06 DCB for MD <50% (42% vs 55-67%), Banks Rm04,R04 significant DCB for non sibling recipient (parent with MD) (17-24% vs 0.5%-2%), Bank T02 with significant (53/261) DCB for future sibling with ID (20% vs 2-5%); 2) Timing of HLA typing: Banks T02,FI03,PV06 with DCB following prenatal HLA (20-27% vs 0%), Banks FI03,Rm04,PV06 with CB HLA typed at birth (61-73% vs 23-25%). Significant HSCT-R (37/177=21 %) was reported by PV06 (HLA selection based storage). Lowest HSCT-R by Rm04,R04 (3/101=3%-5/98=5%) (parental mismatch). At T02 when excluding DCB for future sibling (low birth rate), and autologous (11 ID) HSCT-R increased from 6% (15/261) to 9% (15/170) Conclusion.Advice for public long-term DCB storage should depend on HLA compatibility, potential recipient, disease progression, and likelihood/ timing of using DCB; Italian CB Banks are actively cohoperating to find best public banking practices to ensure a CDB storage system that is ethical, cost effective and responsive to patient needs. Disclosures:No relevant conflicts of interest to declare.

Alternative haematopoietic stem cell sources for transplantation: place of umbilical cord blood.

Umbilical cord blood has rapidly become a valuable alternative stem cell source for allogeneic haematopoietic stem cell transplantation. Extensive research over the last 20 years has established the safety and efficacy of umbilical cord blood transplantation in both children and adults with a variety of malignant and non-malignant diseases. This research has clearly shown that this stem cell source has several unique characteristics resulting in distinct advantages and disadvantages when compared to transplantation with unrelated bone marrow or peripheral blood stem cells. This article reviews the most recent literature comparing the outcomes after umbilical cord blood transplantation with other alternative stem cell sources.

Is mobilized peripheral blood comparable with bone marrow as a source of hematopoietic stem cells for allogeneic transplantation from HLA-identical sibling donors? A case-control study

Granulocyte colony-stimulating factor mobilized peripheral blood stem cells are increasingly used instead of bone marrow as a stem cell source for transplantation. Whereas this change is almost complete for autologous transplantation, there are some concerns when considering allogeneic transplants. We performed a retrospective case-control study including 820 adult patients who had received an allogeneic stem cell transplant from an HLA-identical sibling donor. Quality of life (QoL) was assessed in 150 patients using the EORTC Quality of Life Questionnaire C30 (QLQ-C30). There were no statistically significant differences in overall survival at ten years (bone marrow: 48.9% vs. peripheral blood stem cells: 39.8%; p=0.621), transplant-related mortality (bone marrow: 28.9% vs. peripheral blood stem cells: 34.4%; p=0.682) or relapse incidence at 9 years (29.4% vs. 35.2%, respectively; p=0.688). Similar outcomes were maintained independently of the phase of the disease. However, multivariate analysis identified a higher incidence of acute graft-versus-host disease grades II-IV (p: 0.023; Hazard ratio [HR]: 1.41; 95% confidence interval [CI]: 1.05-1.89) and grades III-IV (p: 0.006; HR: 1.89; 95% CI: 1.20-2.98), in the peripheral blood stem cells-stem cell transplant group. As previously described, extensive chronic graft-versus-host disease was also more frequent in the peripheral blood stem cells group (28% vs. 15.6%; p<0.001). Patients transplanted with peripheral blood stem cells had significant impairment of role and social functioning. Although overall survival was not affected by the stem cell source, peripheral blood stem cell transplants were associated with a higher risk of both acute and chronic GvHD. Global quality of life was similar in both groups, but patients transplanted with peripheral blood stem cells showed worse role and social functioning scores, probably related to the increased incidence of chronic graft-versus-host disease.

Recommendations for a standard UK approach to incorporating umbilical cord blood into clinical transplantation practice: conditioning protocols and donor selection algorithms

Allogeneic haematopoietic cell transplantation is an established curative treatment modality for patients with malignant and non-malignant haematological disorders. Since the first related umbilical cord blood transplant (UCBT) in 1988, the use of UCB as a stem cell source for transplantation has become a standard practice in many countries, with approximately 8000 such transplants having been performed worldwide to date.

제대혈 용기 내부 로봇 암의 열해석에 관한 연구

Umbilical cord blood has been recently considered an attractive potential alternative as a source of stem cell transplantation to curing diseases such as leukemia, cancers, immune disorders. Normally the stored system of the umbilical cord blood specimen is equipped with a computer-controlled robotic arm that enables the samples to locate the identification places in liquid nitrogen tank at regulated temperature as about -196℃. As the half of robotic arm is in the air and the rest part is submerged in liquid nitrogen, the temperature of robotic arm varies from ambient to liquid nitrogen temperature. In this study the temperature variation of upper part of arm above tank lid was thermally analysed by using the commercial code of Ansys. The result of analysis was that the upper part of robotic arm was seriously frozen due to heat transfer from liquid nitrogen as low as -120℃. In order to solve the frost problem of robotic arm, small PTFE tube block as resistance material was introduced into the lower part of tank lid instead of the whole stainless steel(SUS) robotic arm. The results showed that the temperature of robotic arm above the lid was higher enough, and this method would be one of the very effective measure to solve the problem.

Cesarean section due to fetal distress increases the number of stem cells in umbilical cord blood

Umbilical cord blood (UCB) can be used as hematopoietic stem cell source for transplantation. The success of a transplantation is highly correlated with the number of total nucleated cells (TNCs) and CD34+ cells in the UCB. Certain obstetric factors increase the yield of stem cells in the UCB. It is necessary to evaluate optimal conditions in labor to decrease the rate of sample rejection due to low cell count. No data exist regarding the difference between primary and secondary Cesarean sections in terms of efficacy of stem cell harvesting. Seventy-nine consecutive UCB units from women who had a Cesarean section between 1997 and 2003 were included. The number of TNCs, CD34+ cells, colony-forming units (CFUs), white blood cells (WBCs), nucleated red blood cells (NRBCs), and the total collection volume were compared between cases with primary and secondary Cesarean section. UCB obtained after a Cesarean section due to fetal distress has significantly higher numbers of TNCs, CD34+ cells, NRBCs, and WBCs compared to elective Cesarean section. Of the cases with secondary Cesarean section due to fetal distress, 67 percent resulted in UCB units with sufficient TNC numbers (> or =80 x 10(7) TNCs) compared to 42 percent of the cases with primary Cesarean section. Fetal distress increases the number of hematopoietic stem cells mobilized into UCB. Particular effort should be made to collect UCB from newborns who experienced fetal distress.

Synchronized reconstitution of muscle fibers, peripheral nerves and blood vessels by murine skeletal muscle-derived CD34−/45− cells

In order to establish the practical isolation and usage of skeletal muscle-derived stem cells (MDSCs), we determined reconstitution capacity of CD34(-)/CD45(-) (Sk-DN) cells as a candidate somatic stem cell source for transplantation. Sk-DN cells were enzymatically isolated from GFP transgenic mice (C57/BL6N) skeletal muscle and sorted using fluorescence activated cell sorting (FACS), and expanded by collagen gel-based cell culture with bFGF and EGF. The number of Sk-DN cells was small after sorting (2-8 x 10(4)); however, the number increased 10-20 fold (2-16 x 10(5)) after 6 days of expansion culture, and the cells maintained immature state and multipotency, expressing mRNAs for mesodermal and ectodermal cell lineages. Transplantation of expanded Sk-DN cells into the severe muscle damage model (C57/BL6N wild-type) resulted in the synchronized reconstitution of blood vessels, peripheral nerves and muscle fibers following significant recovery of total muscle mass (57%) and contractile function (55%), whereas the non-cell-transplanted control group showed around 20% recovery in both factors. These reconstitution capacities were supported by the intrinsic plasticity of Sk-DN cells that can differentiate into muscular (skeletal muscle), vascular (pericyte, endothelial cell and smooth muscle) and peripheral nerve (Schwann cells and perineurium) cell lineages that was revealed by transplantation to non-muscle tissue (beneath renal capsule) and fluorescence in situ hybridization (FISH) analysis.

Human Cord Blood Cells Transplanted Into Chronically Damaged Liver Exhibit Similar Characteristics to Functional Hepatocytes

Human umbilical cord blood (CB) cells have many advantages as a source for stem cell transplantation because of immaturity and availability. It has been reported that CB cells transplanted into an injured liver displayed hepatocyte-like phenotypes. However, there have been few studies to characterize CB-derived hepatocyte-like cells (HLCs). In this study, CB cells were transplanted into mice with 2 types of liver damage: transient and chronic damage. We analyzed the expression of hepatic differentiation markers in CB-derived HLCs. In the liver of NOD/SCID mice with transient damage, CB-derived HLCs were detected infrequently at 3 weeks after transplantation. In contrast, in the liver of SCID mice damaged chronically by a urokinase-type plasminogen activator transgene under the control of albumin promotor/enhancer (ALB-uPA/SCID mice), more human HLCs colonized the host liver compared with hosts with transiently damaged livers. The CB-derived HLCs in both the transiently and the chronically damaged liver expressed a few markers of human hepatocytes, whereas the transcripts related to mature hepatic functions, including cytochrome P450s, were detected only in the ALB-uPA/SCID mice. These data indicated that CB cells were able to display a similar phenotype to functional hepatocytes in the recipient liver with chronic damage. CB cells may represent a transplantable source for chronic decompensated liver disease.

Four Cases of Donor Cell-Derived Acute Myeloid Leukemia Following Unrelated Cord Blood Transplantation for Adult Patients; Tokyo Cord Blood Bank Experiences.

Donor cell leukemia (DCL) is considered as a rare complication following allogeneic bone marrow transplantation (BMT), whereas the actual frequency of DCL has not yet been specified. Cord blood (CB) is now recognized as an alternative source for stem cell transplantation (SCT), with more than 6000 cord blood transplants (CBTs) performed worldwide, and a few cases of DCL following CBT have also been reported. Here we report four cases of DCL developed after unrelated CBTs using 490 shipped units from Tokyo cord blood bank (TCBB). Development of DCL was informed to TCBB by attending physicians of the recipients in CBT centers as soon as definite diagnosis was made. The feed-back from CBT centers on four DCL cases is summarized in Table 1. All the donors were well at the follow-up questionnaire 6–12 months after birth, but further information of their health conditions has not yet been obtained.Table 1Four cases of DCL following CBTCase1234Recipient32F32F56F30MDiseaseAML in REL1AML-M0 in REL1ATLHodgkin's diseaseSCT1st2nd2nd2ndRegimenMyeloablativeMyeloablativeNon-myeloablativeNon-myeloablativeTBI12Gy12Gy(−)2GyG-CSF(+)(+)(+)(+)GVHD prophylaxisCsA+sMTXCsA+sMTXFK506+PSLCsA+sMTXaGVHDIIII0IIIcGVHD(−)(−)(−)limitedDCLAMLAML-M2AMLAML-M5Latent period15 Mm after CBT11 Mm after CBT7 Mm after CBT16 Mm after CBTDiagnosisFISHSTRFISHSTRThe etiology of DCL is unclear and a common mechanism is unlikely according to the reported literature. There exist several possibilities including occult leukemia or preleukemic state in the donor, defect in immune surveillance, therapy-related stromal abnormalities, excess of cytokine stimulation, and DNA replication and/or repair errors associated with post-transplant expansion of stem/progenitor cells. The possibility of occult leukemia in the donor raises serious problems regarding to the ethical responsibilities of the CBB to the donor.

Incidence and Risk Factors for Hemorrhagic Cystitis in Adult Patients Undergoing Allogeneic Stem-Cell Transplantation.

Introduction: Hemorrhagic cystitis (HC) is a major cause of morbidity in patients submitted to allogeneic stem-cell transplantation (SCT). HC is characterized by a late onset, and it has been related with viral infections, such as cytomegalovirus (CMV), adenovirus, or BK virus (BKV). However, incidence and risk factors for HC are not established so far.Objectives: Analyze incidence and risk factors for HC in a large cohort of adult patients undergoing allogeneic SCT in a single institution.Material and methods: Between 1996 and 2006, 366 adult patients (median age 39 years, range 14 to 63 years) received allogeneic SCT, 251 (69%) from related donor and 115 (31%) from unrelated donor. Source of SCT was peripheral blood (PB) in 219 patients (60%), cord blood (CB) in 95 (26%) and bone marrow (BM) in 52 (14%). Graft was manipulated using CD34+ selection or T-cell depletion in 81 (22%) and 16 patients (4%), respectively. The most frequent underlying diseases were AML (36%), ALL (20%) and CML (15%). Conditioning regimens consisted of busulfan plus cyclophosphamide (BUCY) in 28% of patients, BUCY plus tiotepa (BUCYTT) in 14%, BUCYTT plus thymoglobulin (BUCYTTATG) in 22%, reduced intensity conditioning (RIC) in 12%, and other regimens in 24%. Immunosuppressive therapy consisted of cyclosporine plus methotrexate (52%), cyclosporine plus prednisone (39%) and other (9%). HC was graded according to NCI-CTC modified criteria. In the presence of HC shell-vial urine culture for CMV and adenovirus were performed. Since 2002 VBK viruria using qualitative RT-PCR analysis was also assessed.Results: Overall, 92 patients (25%) developed HC. The median day of presentation was +32 days (range −1 to +680 days). Cystitis cases were graded as follows: grade 1 (4%), grade 2 (40%) and grade 3–4 (56%). Etiology was unknown in 61% of patients, chemotherapy-related in 4% (all from days −1 to +3) and viral-related in 35% (16 VBK, 7 CMV, 6 adenovirus, and 2 HSV). Cumulative incidence (CI) of HC at 3 years was 23%. The following variables were associated with a higher CI of HC: unrelated-donor vs mismatched related donor vs matched related-donor (41% vs 31% vs 16%, p<0.01), CB source vs BM vs PB (47% vs 21% vs 18%, p<0.01), myeloablative regimens vs RIC (29% vs 4%, p<0.01), T-cell depletion vs CD34+ selection (37% vs 17%, p=0.03), BUCYTTATG vs BUCYTT vs BUCY (46% vs 38% vs 13%, p<0.01), age <30 years (41% vs 18%, p<0.01), donor age <30 years (39% vs 14%, p<0.01) and acute GVHD grade 3–4 vs grade 0 (31% vs 15%, p=0.01). In multivariate analysis unrelated-donor and myeloablative regimens were related to a higher CI of HC (both p<0.01). In patients with a matched-related donor CI of HC was higher among these subsets: age <30 years (p<0.01), acute GVHD grade 3–4 vs grade 0 (p=0.02), myeloablative regimens (p=0.02), donor age <30 years (p=0.03), and BUCYTT vs BUCY (p<0.01). In patients with an unrelated donor only age <30 years (p=0.02) was related to a higher CI of HC.Conclusion: In this cohort of adult patients submitted to SCT, the cumulative incidence of HC was 23%. Myeloablative and immunosuppressive conditioning regimens, as well as unrelated-donor, young recipients and young donors are related to an increased risk of HC.