فصلنامه خون
سال ششم شماره 2 (پیاپی 23، تابستان 1388)

Background and ObjectivesMesenchymal stem cells (MSCs) are bone marrow populating cells, which posses an extensive proliferation potential. In isolation and expansion protocols for clinical scale production of MSCs, fetal bovine serum (FBS) is used as a supplement with potential risk for infections as well as immunological reactions. Autologous platelet gel is made from a natural component of the patient’s own blood. Activated platelets release growth factors are mitogenic for MSCs. In vitro studies have indicated that concentration of growth factor varies according to platelet concentration, methods of preparation and mechanism of platelet growth factors release. The aim of our study was to investigate the effect of platelet growth factors on the proliferation and differentiation of human mesenchymal stem cells. Materials and MethodsMononuclear cells of bone marrow were collected in 10% FBS growth medium. The expanded cells were characterized by flow cytometric analysis of specific surface antigens. Analysed markers included CD45, CD34, CD166, CD105, CD90, and CD44. The gel is formed by adding calcium and thrombin to platelet rich plasma (PRP). Treated PRP was incubated for 30 min, 6,24,48 and 72 hours in incubator. Growth factors concentrations in supernatants were determined by ELISA. Human mesenchymal stem cells were cultured in the complete medium that supplemented with 10% FBS or Platelet growth factors for 8 days. The rate of proliferation was evaluated by MTT assay. Expanded cells were seeded on calcium phosphate scaffold. Cells growth and morphology on scaffold were analyzed by SEM. ResultsIsolation and expansion of MSCs in the complete medium supplemented with platelet growth factors were successful and morphology of cells was compatible with that of FBS. Cells were highly positive for CD90, CD166, CD44 and CD105 and negative for CD34, CD45. There was no significant difference between expression of markers on cells expanded with platelet growth factors and FBS. We demonstrated that platelet growth factors provide a significantly higher proliferative effect on MSCs than those of FBS. MSCs cultured in the presence of growth factors maintain their osteogenic differentiation properties. Osteogenic differentiation was indicated by deposition of mineralized matrix stained with Alizarin red and increased expression alkaline phosphates. ConclusionsPlatelet growth factors can be used in place of FBS to provide a safer and more effective culture condition to expand MSC for clinical purposes. MSCs cultured in the presence of platelet growth factors maintain their osteogenic properties.

Background and objectivesOne of the major etiologies of lipid disorders in malignancies is the use of chemotherapy drugs, the most important of which is L-Asparginase. Studies in different centers with high dosages of L-Asparginase demonstrated different results. The aim of this study is to evaluate the effect of L-Asparginase at a dose of 6000 U/m2 on lipid profile in Iranian children with newly diagnosed acute lymphoblastic leukemia in Mofid Children Hospital of Tehran. Materials and MethodsWe performed a nonrandomized trial in which all children with newly diagnosed acute lymphoblastic leukemia (ALL) aged <15 years participated. Every case serves as his/her own control. The cholesterol, triglyceride (TG), HDL, LDL, VLDL, APOA, APOB and LPa were evaluated in three stages: before, during and 2 months after treatment with L-Asparginase after which the results were compared. ResultsIn our study, 82 newly ALL diagnosed patients with the mean age of 6 years (within the age range of 0.12-14 and SD of 3.5) were evaluated. Mean seum level of TG in pretreatment stage was 163.9 mg% and during treatment with L-Asparginase 123 mg% demonstrating significant decline in TG serum level after administration of L-Asparginase (p=0.002). Mean serum level of cholesterol before administration of L-Asparginase was 151mg% and during treatment 140 showing no statistically significant difference (p= 0.061). LPa level in pretreatment phase was 24mg% and during treatment 14 showing a statistically significant difference (p= 0.0001). ConclusionsL-Aspar resulted in decline in TG serum level and increase in HDL though with no significant difference in cholesterol level. Overall, L-Asparginase even at a dose of 6000U/M2 does not raise blood level of triglyceride and cholesterol.

Background and Objectives Hematopoietic stem cells (HSCs) transplantation has been used for treating hematological disorders, immunodeficiencies, metabolic disorders and auto immune disease. In clinical application, the number of HSCs infused proved to be the major prognostic factor for engraftment and survival; however, cord blood transplantation is limited in adults cause of their low number of cells. Consequently, we evaluated the potential of cell expansion and differentiation of cord blood cells in presence of different combination of cytokines. Materials and MethodsIn this interventional experimental study, umblical cord blood mononuclear cells (UCB MNC) were taken from apparently healthy mothers without any historical sign of diabetes during their pregnancy. We quantified and characterized an ex vivo expansion capacity of umbilical cord blood mononuclear cells (UCB MNC) in IMDM supplemented with 10% fetal bovine serum and various combination of SCF, TPO, Flt-3, GM-CSF and IL-3 as FSG3 T, FS3T, and FST for 3 weeks. Immunophenotyping and colony forming assay were performed at day 0, 7, 14 and 21 to characterize expanded cells in different groups. ResultsTotal cell number was multiplied 5 times by all groups after 14 days of culture and the group with three cytokines of SCF, TPO, Flt-3 was shown to produce the highest percentage of hematopoitic stem cells and myeloid precursor cells among all groups during 2 weeks. ConclusionsIn this study, we showed an economical method for the expansion of umbilical cord blood stem cells with usage of UCB MNC and combination of SCF, TPO, and Flt-3.

Background and Objectives Blood typing by serologic methods after transfusion has limitations due to presence of donor red cells in recipients. Accurate determination of red blood cells (RBCs) antigens is very important in multitransfused patients including beta-thalassemics and sickle cell anemics. So, the aim of this study was to evaluate DNA- based methods as supplement to the hemagglutination technique to determine the red blood cell (RBC) antigen profile of multitransfused patients with beta- thalassemia. Materials and Methods DNA was extracted from peripheral blood of 20 apparently normal people and 44 patients including 35 with beta- thalassemia (out of whom 19 had clinical evidence of delayed hemolytic transfusion reaction), 8 with thalassemia intermedia (out of whom 2 had hemolytic reaction), and one with sickle cell thalassemia. RHD/ RHC/ RHc/ RHE/ RHe/ JKA/ JKB/ FYA/ FYB/ KELL1/ KELL2 alleles were determined by PCR and were then compared with the hemagglutination method. Results Phenotype and genotype results were the same in all controls. The phenotypes and genotypes of 53 blood antigens of 26 patients were incompatible. Most of the discrepancies (19 cases) occurred in the Rh system, and fifteen in the Duffy and Kidd systems.Conclusions The results show that screening platelet concentrates for bacterial contamination is necessary for blood transfusion centers and hospital blood banks. Blood typing by serologic method was not accurate in this study but genotyping could determine true blood groups in multitransfused patients and help in selection of RBCs without alloimmunized antigens in future transfusion attempts. Specificity, sensitivity, positive and negative predictive values of hemagglutination method for RhD antigen had good values in comparison to the molecular method. This might be due to pre- transfusion determination of RhD for thalassemic patients so as to receive Rh- matched blood units. It seems pre-transfusion blood typing of Rh and Kell antigens, which are the cause of hemolytic reactions, in comparison to the molecular method could be cost effective. In addition, typing of Rh and Kell antigens in some regular blood donors could be helpdul for selecting antigen-negative RBCs for transfusion dependent patients

Background and ObjectivesTransfusion transmitted HIV is an important issue in every transfusion center. Despite recent improvements in laboratory screening techniques and formulation of rigid standards for blood safety, the risk of HIVcontaminated blood products remains as a potential problem. This study was conducted to determine the knowledge, attitude and practice regarding blood donation among voluntary blood donors about HIV/AIDS. Materials and MethodsThis is a cross-sectional study in which data were collected through questionnaires.We studied knowledge, attitude and behavior of 440 voluntary blood donors about HIV/AIDS in Sistan and Balouchestan province. Voluntary blood donor's knowledge and attitude were evaluated and their correlation with different variables were compared. The data were analyzed with Chi-Square, ANOVA, Kruscal Wallis and Spearmans rho using SPSS 14. ResultsOur results showed that the knowledge levels of 55% of voluntary blood donors about HIV/AIDS were good, 29.5% intermediate, and 15.5% poor. Seventy point one percent of population in the study had positive attitude,56.9% had neutral attitude and 25.9% negative attitude about HIV/AIDS. The correlation between sex, marriage status, educational status, job, type of donation (voluntary or familial),times of donation (first, repeated and regular repeated) with knowledge and attitude was statistically significant. ConclusionsRelatively high level of knowledge about HIV/AIDS especially in regular repeated donors can be due to acquired information during consultation sessions. Therefore, repeated and regular repeated donors can help improve the blood safety.These findings also highlight the importance of more accurate consultation and the provision of necessary training on HIV/AIDS to voluntary blood donors.

Background and ObjectivesMany of burn patients need to receive blood transfusion during debridement and skin grafting. Although it is vital, blood transfusion adverse effects and risks are not unlikely. Therefore, it is better to minimize blood transfusion. In this study, we have tired to estimate necessary blood volume requirement in proportion to preoperative patient hematocrite level and burn percentage that would eventually lower inappropriate blood use. Materials and MethodsThis prospective study was done on all patients undergoing debridement and grafting skin surgery through 6 months. Demographic data, burn percentage, reserved and transfused blood, preoperative and postoperative Hct level were recorded. Qualified data were analyzed with Chi - square and regression analysis (p-value<0.05). ResultsMean age of all patients was 23 with burns affecting 33 ± 21% of total body surface area. For all patients 181 blood units were reserved and 85 were transfused. But if indication for transfusion was determined to be Hct ≥ 28%, blood requirement would be just 54 units for 33 patients. "Requirement of blood transfusion" had negative correlation with "preoperative Hct" (p = 0.001), and positive correlation with "burn percentage" (p = 0.009). ConclusionsThe blood volume requirement for debridement and skin graft surgery in burn patients can be estimated by "preoperative Hct level" and "burn percentage"; this process can minimize unneccessary reserved and transfused blood units significantly.

Background and ObjectivesAlthough major beta-thalassemia prevention and control program is one of the most successful programs in genetics field, there are new cases of major beta-thalassemia due to laboratory errors. So the laboratory quality control (QC) is very important to diagnose carriers in preventive programs. At present, there is not a distinct and measurable index for function assessment and comparison in screening laboratories; this urged us to create an objective, measurable and explicit index for health care givers. For this purpose, we compared the function of screening labs in districts of Isfahan Province. Materials and MethodsIn this cross-sectional study, we collected the data from "pre-marriage screening program" in 18 districts (19 screening labs) covered by Isfahan University of Medical Sciences from 2006 to 2008. The percentage rates of "screened males with low blood indexes" in every 3 months (P index) were assessed; then, the average and coefficient variants in duration of 3 years were calculated with excel software. Based on these findings, "screening labs’ functions" were assessed and compared. ResultsThe least "CVs of P index" within 19 labs were found to be in Mobarakeh laboratory (10.4%) and Provincial reference lab (10.9%), respectively. The highest CVs were found in Khour (76.3%) and Tiran and Karvan labs (58.2%), respectively. The P index average in all centers was 14% (13.9% in reference lab) with the highest being in Mobarakeh(24.2%) and the lowest in Khour(8.8%). ConclusionsWhereas the number of marriage candidates in each screening lab is relatively stable, the centers with the highest CV value of P index are probably lower in function level as compared with the centers with the lowest CV value of P index (as in reference screening lab). CV of P index may be a good evidence for quality of "screening lab function" and should be evaluated annually. The screening labs with high CV of P index need more attention and evaluation.

Background and ObjectivesBlood reservation is done in many surgical operations because of the risk of bleeding. However, reservation should be based on a standard protocol so as to prevent blood loss. In this study, we deal with the ratio and causes of unused blood ordered from blood bank in elective surgeries in Rasoul-e-Akram hospital in Tehran. Materials and MethodsIn this descriptive study, a questionnaire was added to file records of patients who were candidates for elective surgery in Rasoul-e Akram Hospital during 2005. Questionnaires were filled out by the physicians who had ordered blood after patient's discharge; these questionnaires were ultimately sent to blood transfusion committee. The collected data were then analyzed using descriptive statistic methods and SPSS version11.5. ResultsThe total number of reserved blood bags for elective surgeries was 8385 in one year out of which 1460 (17/4%) were transfused. The ratio of transfused cross-matched blood was 3.12; the major reason for ordered blood not to be used was incorrect surgical team estimation (44.6%). ConclusionsThis study revealed that blood reservation is not performed based on any definite protocol and it is necessary to control the reservation and use of blood both qualitatively and quantitatively in medical centers.

Blood-letting is defined to be the withdrawal of blood from a patient. Considering the mysterious, life-saving, and occassionally miraculous nature of blood during the evolving history of man, civilization, and science, this red liquid being the token of life and death throughout centuries was used as evidence for clinical diagnosis of special diseases or otherwise as definite and soothing treatment of patients. Based on the existing evidence, hijama or blood withdrawal in cultural and religious beliefs and customs of certain tribes has had even a special status in saving man from devil or evil forces. The accessible old documents show the expansion of blood drawing as a known life-saving element and treatment method in ancient Egypt, Greece, and Rome in different forms including hijama. However, the development of medical sciences, particularly transfusion medicine and blood transfusion sciences, the treatment and preventive role of hijama and other methods like arteriotomy and leech cupping started to get less prominence except for some eastern countries especially Islamic states where hijama is still employed to relieve soul and treat diseases as a tool of preserving traditions. In Iran, considering the available standards based on which potential blood donors with recent hijama experience are defered for one year, it is necessary to raise awareness of all those involved in the field of blood transfusion and the whole community about the history of hijama so as to see how we can better deal with this historical and traditional controversial topic.