Showing 14 results for Jalili
H Khorsand Mohammad Pour, I Nourmohammadi, M.a Jalili, Z Motallebi,
Volume 3, Issue 1 (Spring 2006)
Abstract
Abstract
Background and Objectives
Albumin is currently used in greater volume than any other biopharmaceutical solution that is available. The most large- scale production of human albumin is still conducted by cohn cold ethanol fractionation (Cohn's method). In order to save the operating time, labor and material in manufacturing albumin 20%, a two-stage process was carried out based on the cold ethanol fractionation method.
Materials and Methods
For this experimental study, fresh frozen plasma (FFP) was used as a starting material. In the first stage by addition of supernatant IV to a dissolved fraction V paste, a dense fraction V paste (Vd+I) containing the highest possible albumin concentration and the least impurity (ethanol, a globulins and ionic concentration) was recovered by centrifugation the recovered paste was then dissolved and filtered through a depth filter in order to remove a globulins as impurity. The filtrate (HPs) was suitable to produce albumin 20%. Two batches were produced and all the methods for the evaluation of the finished product were applied according to the guidelines of the British Pharmacopoeia.
Results
The quality control of the albumin 20% as a finished product from 2 batches (batches 1 and 2) in the final container produced by the proposed procedure led to satisfactory results. Appearance of the product was clear, purity over 99% , and the molecular size distribution of albumin was revealed for aggregates or polymers less than 2.7 % . The yield of albumin was 24 ± 1 gr/kg of plasma.
Conclusions
Advantages of the proposed process as compared with the rountine process (Cohn & Kistler- Nitschmann methods) during the production of intermediate products (Vd+I, HPs) were remarkable savings in terms of operating time, material and manpower at about 50%.
Key words: Human serum albumin, Plasma fractionation, Cohn Fraction V
A Aghaie, A.a Pourfathollah, S.z Bathaie, S.m Moazzeni, H Khorsand Mohamad Pour, H Rezvan, S Banazadeh, B Adibi, M.k Mosavi, M.a Jalili,
Volume 3, Issue 2 (Summer 2006)
Abstract
Abstract
Background and Objectives
Human plasma is a valuable and unique material containing vital proteins such as albumin, immunoglobulin, and coagulation factors which have pharmaceutical applications. The most current process for purification of immunoglobulin is the Cohn fractionation procedure using ethanol. The basic material for production of immunoglobulin in this procedure is fraction II. In the present study, Cohn fractionation was modified appropriately in order to achieve fraction II with a quality suitable for production of intravenous immunoglobulin.
Materials and Methods
Various plasma proteins were precipitated using varying concentrations of ethanol under controlled physiochemical conditions such as temperature , pH and ionic strength. The fractionation products as well as the resulting fraction II were tested and analyzed.
Results
The results demonstrate that the modified procedure used can result in a pure fraction II with high yield . The procedure was also suitable for all other plasma proteins. The purified fraction II contained acceptable PKA levels according to specifications of pharmacopoeia. The protein structure was also within normal limits. The repeatability of the modified procedure reported was acceptable.
Conclusions
The fraction II obtained in the modified Cohn procedure was a suitable intermediate product to be used in production of intravenous immunoglobulin . The PKA levels as well as the protein aggregation were minimal , and the quality of fraction II was standard . The results showed that the present modified Cohn procedure can be easily scaled up to industrial and semi-industrial levels. The resulting fraction II can be used as an intermediate in production of IVIg.
Key words: Intravenous immunoglobulin, Cohn fraction, Molecular conformation
Dr Mehryar Habibi Roudkenar, R Halabian, N Shagerdi Esmaili, A Oodi, N Masroori, Dr N Amirizadeh, Dr K Mousavi Hosseini, Dr A Gharehbaghian, Dr H Rezvan, Dr M.a Jalili,
Volume 6, Issue 1 (Spring 2009)
Abstract
Isolation, cloning and expression of recombinant
human factor VII in CHO cell line
Halabian R.1( MS ), Shagerdi Esmaili N.1( MS ), Oodi A.1( MS ), Masroori N.1(MS), Amirizadeh N.1(PhD), Mousavi Hosseini K.1(PhD), Gharehbaghian A.1(PhD), Rezvan H.1(PhD), Jalili M.A.1(PhD),
Habibi Rudkenar M.1(PhD)
1Iranian Blood Transfusion Organization, Research Center, Tehran, Iran
Abstract
Background and Objectives
Factor VII is a plasma glycoprotein that participates in the coagulation process leading to the generation of fibrin. Factor VII plays an important role in the cascade of coagulation. The aim of this study was to clone and express human recombinant factor VII in CHO cell line as an eukaryotic host cell.
Materials and Methods
In this descriptive study, FVII cDNA was isolated from HepG2 cell line and cloned to pcDNA 30.1(+) vector. The constructs were transfected to CHO cell line. A cell line that permanently expressed recombinant factor VII was established. The expression of recombinant FVII was determined by RT-PCR, ELISA, SDS-PAGE and western blot analysis. Biological activity of recombinant factor VII was determined by prothrombin time assay in factor FVII-depleted plasma.
Results
The results showed that FVII was successfully cloned and expressed. After 3 weeks stable cell lines were generated in the culture of the CHO cell line in the presence of geneticin. RT-PCR, ELISA, SDS-PAGE and western blot analysis results indicate the expression of FVII in the stable clones. A three- to four-fold decrease of the specific coagulant activity of rFVII was observed that indicates of rFVII being biologically active.
Conclusions
Four thousands to Six thousands vials of rFVII were imported to our country having high cost for the government. Therefore, production of rFVII through recombinant DNA technology within lab scale is the first step to overcome the problems.
Key words: Hemophilia, rFVIIa, CHO, Transfection
SJIBTO 2009 6(1): 1-11
Received: 21 Oct 2008
Accepted:21 May 2009
Correspondence: Habibi Roudkenar M., PhD of Biotechnology. Assistant professor of Iranian Blood Transfusion Organization-Research Center. P.O.Box: 14665-1157, Tehran, Iran. Tel: (+9821)88601599 Fax: (+9821)88601599
E-mail: roudkenar@ibto.ir
A. Talebian, A.r. Shafaei, M. Sharafi, S. Rivandi, L. Jalili, T. Fattaheian,
Volume 7, Issue 1 (Spring 2010)
Abstract
Abstract
Background and Objectives
ABO blood group system antibodies agglutinate red blood cell suspension in physiologic serum directly without any reaction booster. In the present study the potency of IBRF-manufactured anti-A and anti-B blood grouping reagents by the hemagglutination method in test tube was reassessed against the new WHO international minimum potency standards.
Materials and Methods
In this experimental study, starting concentrations defined in WHO standards were used in titration. Doubled dilution series of WHO standards (from starting concentrations) and IBRF blood grouping reagents (from neat) were prepared by using buffered saline containing 2% BSA as diluent. One volume of each starting concentration together with one volume of prepared dilutions were mixed with one volume of a 2% suspension of A1, A2, A2B, and B cells in glass test tubes, respectively. After appropriate incubation and centrifugation of the tests according to specified criteria, the reactions were graded macroscopically.
Results
The results showed that the IBRF anti-A and anti-B blood grouping reagents comply with the minimum WHO standard dilution. Consequently, IBRF anti-A and anti-B blood grouping reagents were shown to be safe for screening and diagnostic purposes.
Conclusions
The quality of blood grouping reagents is clearly an important factor for safe blood transfusion. Routine titration tests are not reliable methods for evaluation of those reagents. Fortunately, this problem would be solved using the above standards. This recommended method is provided to help assist manufacturers in pursuing new product license applications and making amendments in existing ones.
Key words : Reagents, Agglutination, Standards
M. Ashki, Dr. N. Amirizadeh, Dr. M.a. Jalili, Dr. N. Hayati Roudbari, M.h. Mohammadi, M. Amani,
Volume 8, Issue 4 (Winter 2012)
Abstract
Abstract
Background and Objectives
During differentiation of mesenchymal stem cells (MSCs) into various cells, the expression of a variety of genes undergoes some changes in this study we decided to investigate the expression rate of some genes like osteopontin (OPN) and osteocalcin (OCN) during this process in order to find a better and faster way for these cells to be differentiated into osteoblasts .
Material and Methods
In this experimental study, the mononuclear cells of bone marrow were separated and then cultured in DMEM-LG culture media with 10% FBS. During some definite days, the RNA of differentiating cells was extracted. Then, the effective genes in osteogenesis like OPN and OCN were amplified by speciefic primers. The mesenchymal cells were cultured on 3D calcium phosphate scaffolds, and finally the activity rate of the alkaline phosphatase was examined .
Results
This research has demonstrated that in the process of differentiation, the expression of the two genes of OPN and OCN changed orderly with the maximum expression of OPN in the 6th day and the maximum expression of OCN in the 7th and 8th days of differentiation. The osteogenic differentiation of MSCs was not confirmed by the coloration of mineral sediments. The activity rate of alkaline phosphatase revealed the preference of 3D calcium phosphate scaffold to 2D environment in this differentiation.
Conclusion
The calcium phosphate scaffold positively affects the differentiation process. The expression of OPN and OCN genes changes during differentiation and can be used as away to a better and faster differentiation of these cells into osteoblast.
P. Hamedi Asl, R. Halabian, M. Mohammadzadeh, M. Mohammadipour, Z. Bakhshandeh, Dr. Hamedi Asl, A.a. Kianin, Dr. M.a. Jalili, Dr. N. Amirizadeh, Dr. M. Habibi Roudkenar,
Volume 9, Issue 3 (Stem Cell Supplement , Autumn 2012)
Abstract
Abstract
Background and Objectives
Heme oxegenase1 (HO-1) is one of the potent cytoprotective factors. The goal of this study was to perform cloning and transient over expression of the human HO-1 gene in mesenchymal stem cells (MSCs) using the adenoviral expression system based on the gateway technology.
Materials and Methods
In order to induce expression of HO-1, A549 cell lines were exposed to UV for 1 hour. The full length cDNA of HO-1 was isolated and cloned into pENTR TOPO/D vector by TOPO cloning reaction. To construct the expression clone, a LR recombination reaction was carried out between the entry clone and destination vector, pAd/CMV/V5-DEST. The recombinant virus was produced in the appropriate mammalian cell line. MSCs were infected by the recombinant virus expressing HO-1.
Results
The results showed that human recombinant HO-1 was successfully cloned and the accuracy of the gene and its frame in the vector were confirmed by DNA sequencing. Expression of HO-1 in MSCs was confirmed by RT-PCR and western blot analysis. The results indicated that the expression of HO-1 is transient.
Conclusions
Transient expression of human HO-1 gene in MSCs by using adenovirus expression system may be considered as an efficient gene transfer strategy into MSCs in order to promote stem cell therapy.
A. Hosseini, R. Halabian, Dr. P. Hamedi Asl, H. Bashiri Nahanji, Dr. M.a. Jalili, M. Heydari, Dr. N. Amirizadeh, Dr. M. Habibi Roudkenar,
Volume 10, Issue 1 (Spring 2013)
Abstract
Abstract
Background and Objectives
Previous studies have showed that a large percentage of Mesenchymal Stem Cells (MSCs) die in the early stages of transplantation due to oxidative streeses, hypoxia, and serum deprivation. Hence, this study was aimed to address whether induction or inhibition of autophagy would affect the viability of MSCs after exposure to oxidative stress.
Materials and Methods
In this descriptive study, MSCs were isolated from bone marrow with ficol and density gradient method the fourth passage MSCs were selected in the study. Autophagy was detected by using transfection of GFP-LC3 to MSCs. Induction and inhibition of autophagy were performed by using rapamycin and 3MA, respectively. MSCs were exposed to lethal doses of H2O2 followed by cells viability evaluated with MTT assay.
Results
Our results revealed that the enhancement of autophagy in MSCs sensitized them against oxidative stress and inhibition of autophagy led to resistance against oxidative stress in comparison with the control cells.
Conclusions
Inhibition of autophagy using non genetic engineering method in MSCs enhances cell viability following exposure to the oxidative stress. This may provide a novel strategy to promote the efficiency of cell therapies following transplantation in the future.
Key words: Mesenchymal Stem Cells, Autophagy, Oxidative Stress, Cell Hypoxia, Rapamycin
Dr. T. Chahkandi, Dr. T. Kazemi, Dr. A. Jalili, Dr. F. Ghaderi,
Volume 10, Issue 1 (Spring 2013)
Abstract
Abstract
Background and Objectives
The most common leading causes of death in patients with major beta thalassemia are cardiac complications.The purpose of this study is to determine cardiac function in patients with major thalassemia in Birjand.
Materials and Methods
This descriptive-analytical study was conducted on all patients with major beta thalassemia having referred to special clinics of Valiasr Hospital in Birjand. Variables such as age, sex, frequency of transfusions, serum ferritin, and hemoglobin were studied. For all patients after a thorough examination of heart, ECG, chest X-ray, and echocardiography were performed. All data were collected and analyzed with SPSS software 15.5 by t-test at α ≤ 0.05.
Results
We studied 35 patients with major beta thalassemia. The mean age was 9.06 ± 4.33 years. The mean hemoglobin was 9.2 gr/dl and the mean ferritin 2121.6 ng/ml, respectively. Cardiac examinations showed 62.9% of patients to be normal. ECG and chest X-ray in 71.6% and 57.1% of patients were normal, respectively. Out of the total number of patients, 77.8% had abnormal echocardiography. The most common abnormal findings in echocardiography were restrictive diastolic dysfunction. No significant correlation was found between abnormal findings on echocardiography with both ferritin and the number of blood transfusions. However, left ventricular diastolic dysfunction and hemoglobin were not significantly associated (8.5 ± 0.69 gr/dL in normal diastolic function, 9.6 ± 0.99 gr/dL in diastolic dysfunction p= 0.007).
Conclusions
The most common finding in our study was left ventricular diastolic dysfunction. Therefore, periodic echocardiography to determine the risk of heart involvement is very valuable.
H. Bashiri Nahanji, Dr. M. Habibi Roudkenar, R. Halabian, Dr. M. Jalili, Dr. M.a. Jalili,
Volume 10, Issue 3 (Autumn 2013)
Abstract
Abstract
Background and Objectives
About 70% of MSCs die in the early stages of transplantation into the infarcted myocardium. Several solutions have been made to address this problem. In the last decade, preconditioning of MSCs with oxidative stresses has gained a lot of attention. In this study, we have investigated the effects of preconditioning with hydrogen peroxide (H2O2) on the survival of MSCs and their resistance against oxidative stresses .
Materials and Methods
Mesenchymal stem cells from bone marrow have been cultured. Cells from the passage four were treated with 5, 10, 15, 20, 30, 40, 50, 60, 80, and 100µm concentrations of H2O2, and were then recovered with the fresh medium. Finally, the treated cells were exposed to 500 µM H2O2 as the killing condition. The percentage of survived cells was analyzed by the MTT assay kit .
Results
Preconditioning with 5 and 10 µ M H2O2 significantly increased the resistance of MSCs against the apoptosis induced by 500 µ M H2O2 .
Conclusions
Preconditioning of MSCs with oxidative stresses enhances their survival therefore, it can increase the efficacy of transplantation .
F. Nasiri, Dr. F. Amiri, M. Mohammadipour, S. Molaei, Dr. M. Habibi Roudkenar, Dr. M.a. Jalili,
Volume 12, Issue 2 (Summer 2015)
Abstract
Abstract
Background and Objectives
Mesenchymal stem cells (MSCs) are attractive cells for cell therapy. However, the efficacy of MSCs is limited because of low survival rate following translation. In this study, MSCs were preconditioned with sub-lethal doses of H2O2. The therapeutic potential of preconditioning MSCs was examined on acute liver failure in mice.
Materials and Methods
In the present experimental study, MSCs were isolated from bone marrow aspirate. Cells from passage four were treated with different concentrations of H2O2 for 24 hrs, and then were recovered in fresh medium for 7 hrs followed by exposure to lethal doses of H2O2. Cell viability was measured with WST-1 assay. In the in vivo phase, acute liver failure was induced by CCl4 then, the regenerative potential of preconditioned-MSCs was evaluated using biochemical as well as histological methods. |
Results
Survival rates were higher in the engrafted mice with preconditioned-MSCs in comparison to those who received normal MSCs. Three days after cell therapy liver enzymes reached the normal level in engrafted group with preconditioned-MSCs. Interestingly, histological results revealed a significant improvement in liver regeneration potential in transplanted groups with preconditioned-MSCs.
Conclusions
Preconditioning of MSCs with H2O2 not only enhances their survival but also increases the efficacy of MSC-based cell therapy in liver acute failure.
H. Mehrabi Habibabadi, Dr. F. Amiri, Dr. E. Moslemi, Dr. M. Habibi Roudkenar, M.a. Jalili,
Volume 12, Issue 3 (Autumn 2015)
Abstract
Abstract
Background and Objectives
Due to the unique criteria, mesenchymal stem cells (MSCs) are the ideal cells for cell therapy and gene therapy. However, the low survival of MSCs after transplantation has limited their application. This study aimed to evaluate the expression of cytoprotective genes including NQO1, TXNRD1, HO-1, GCLC following the overexpression of Nrf2 in MSCs.
Materials and Methods
In this experimental study, umbilical cord-derived MSCs were cultured and recombinant vectors containing Nrf2 and empty vectors were transfected into MSCs using FuGENE HD. After exposure of the cells to stress, RNA extraction and cDNA generation were performed. Using Primer3 software, specific primers were designed for Nrf2, NQO1 ، TXNRD1 ، HO-1 and GCLC genes and the expression of these mentioned genes was evaluated by RT-PCR. The results were quantified and analyzed statistically utilizing Image J software and ANOVA. |
Results
The expression of Nrf2 was up-regulated in MSCs after transfection (p< 0.01). Overexpression of TXNRD1 and GCLC was observed in transfected cells (p <0.05 and p <0.01) however, the expression of NQO1 and HO-1 did not change in the transfected group in comparison to the control (p > 0.05).
Conclusions
Overexpression of Nrf2 resulted in the overexpression of TXNRD1 and GCLC in MSCs and might be explained by the fact that a part of the known Nrf2 cytoprotective mechanisms is controlled by the expression of these genes.
F. Soltani, Dr. F. Amiri, Dr. M. Mohammadipour, M. Jalili, Dr. M. Habibi Roudkenar, Dr. M.a. Jalili,
Volume 12, Issue 4 (Winter 2016)
Abstract
Abstract
Background and Objectives
Mesenchymal Stem Cells (MSCs) are the ideal cell source for transplantation. But different stresses during MSCs in vitro expansion lead to decreased survival rate after transplantation. Therefore, applying practical strategies to enhance their viability in stressful microenvironment is quite necessary. This study aimed to survey effects of HIF-1&alpha-Nrf2-engineered-MSCs secretome on MSCs survival under different stress conditions.
Materials and Methods
|
Recombinant pcDNA3.1-Nrf2 and pcDNA3.1-HIF-1&alpha were transfected and co-transfected into umbilical cord MSCs (UC-MSCs) using FUGENE HD transfection reagent. After 72 hrs, expression of Nrf2 and HIF-1&alpha were verified by RT-PCR. Different cell groups were exposed to hypoxic, serum deprived and oxidative stress conditions. The HIF-1&alpha-Nrf2-engineered-MSCs secretome was harvested and concentrated. Then, UC-MSCs were cultured in presence of this secretome and their viability was assayed using trypan blue exclusion dye and WST-1 flowing by induction of the same stress conditions.
|
Results
HIF-1&alpha-Nrf2-engineered-MSCs expressed Nrf2 and HIF-1&alpha. HIF-1&alpha-Nrf2-engineered-MSCs indicated a higher survival rate (84.5 ± 5.5%) compared with the control group (55.3 ± 4%). Moreover, the survival rate of UC-MSCs cultured with HIF-1&alpha-Nrf2-engineered-MSCs secretome was 81.6 ± 6% and it was 57.9 ± 4.3% for the control group under the same stress conditions.
Conclusions
HIF-1&alpha-Nrf2-engineered-MSCs secretome protects MSCs against oxidative, serum deprived and hypoxic stress conditions.
F. Jaleh, F. Amiri, M. Dehghan Harati, Dr. M. Habibi Roudkenar, Dr. M.a. Jalili,
Volume 13, Issue 4 (Winter 2016)
Abstract
Abstract
Background and Objectives
The decrease in mesenchymal stem cells (MSCs) survival rate after transplantation is a major challenge in MSC-cell-therapy. Hence, it is promising to employ some proper strategies for addressing this problem. This study was conducted to assay the therapeutic effects of MSCs treated with Nrf2-manipulated-MSC conditioned medium in acute kidney injury (AKI)-induced animal models.
Materials and Methods
|
In an experimental study, recombinant plasmid pcDNA3.1-Nrf2 was transfected into bone marrow-derived MSCs and the resulted conditioned medium was harvested. The MSCs was cultivated with Nrf2-manipulated-derived conditioned medium. The conditioned medium-treated-MSCs were transplanted to AKI-induced rats (each group containing 10 rats) and the therapeutic potentialities of these MSCs were evaluated using biochemical and pathological methods.
|
Results
Fourteen days after MSCs transplantation, there was a significant difference in BUN decreasing in rat groups injected with conditioned medium-treated-MSCs (48 ± 3.5 mg/dL) in comparison with those injected with normal MSCs (100 ± 9.9 mg/dL) (p < 0.001). The number of observed casts (0.26) in kidney tissue sections of these rat groups were also less than (0.51) the groups transplanted with normal MSCS.
Conclusions
Cultivation of MSCs in the presence of Nrf2-manipulated-derived conditioned medium increase the therapeutic effects of these cells in AKI-induced models.
M. Nayebhashemi, M. Mohammadi Pour, H. Fahimii, M. Habibi Roudkenar, Mohammad Ali Jalili,
Volume 14, Issue 1 (Spring 2017)
Abstract
Abstract
Background and Objectives
Stem cell factor (SCF) is a 28-40 kDa glycoprotein which plays an important role for the proliferation and differentiation of hematopoietic stem cells (HSCs). SCF binds to the C-kit receptor, and improves the survival of HSCs in vitro. SCF is one of the essential supplements for cultivation of HSCs in vitro. It plays a vital role to increase the number and size of HSC colonies.
Materials and Methods
|
In this experimental study, glycosylation of SCF does not seem to influence its biological activity; therefore, it would be possible to produce it in prokaryotic expression system. The aim of this study was isolation, cloning and expression of SCF in the Rosetta expression host. In addition to the features of prokaryotic expression system, Rosetta was expected to provide rare codons of eukaryotic proteins and increase the recombinant protein expression level.
|
Results
The SCF coding sequence was isolated and amplified using the specific primers and cloned in E.coli TOP10 using pET-32a expression vector. The recombinant construct was confirmed by PCR, digestion and sequencing. The recombinant vector was transformed to the expression host, Rosetta.
Conclusions
The SCF coding sequence was successfully isolated and cloned into the expression vector pET-32a, followed by recombinant protein expression in Rosetta host strain.
