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Showing 9 results for Culture

Golestani R, Moazzeni S. M, Puorfathollah A.a, Hamidpour L, Sharafi M, Attarchi Z, Hadji-Beygi B, Tavasoli F, Esmaili J, Reyvandi S,
Volume 3, Issue 3 (10-2006)
Abstract

Abstract

Background and Objectives

Providing fetal calf serum (FCS) alternatives as cell culture supplements is an important field of research to compensate for the FCS supply shortage.This study focused on preparation of fetal calf serum alternatives and their effects on growth and secretion of hybridoma cell lines.

Materials and Methods

Outdated human platelet units undergo extraction for its growth factors to be obtained. Human AB blood group plasma was also converted to serum and its growth effect was compared to FCS, hypoxanthine-thymidine (HT) and RPMI1640 as cell culture media and supplement. Cell growth indices were preliminary counting of cells , confluency as surface area of plates filled with cells, and titration of monoclonal anti-A and anti-B blood group antibodies collected from cultured mouse hybridoma cells. Statistical analysis including one sample t-test, logarithmic multiple regression curve fit, and factor analysis was done by SPSS v12 software.

Results

The four nutritional supplements of (1) human serum AB (AB), (2) human platelet extract (PLT), (3) equal mixture of AB & PLT (ABP), and (4) fetal calf serum as cell culture were examined on mouse hybridoma anti-A and anti-B monoclonal antibody producer cell lines for cell growth indices and compared with the same indices on RPMI1640 media. The growth-stimulating effects in descending order of values were (1) ABP5% , (2) FCS10% , (3) ABP10% , (4) AB5% , (5) AB10% , (6) PLT5% , (7) ABP20% , (8) PLT10% , (9) PLT20%, and (10)HT but AB20% inhibited growth of mentioned hybridoma cell lines. The titer of anti-A and anti-B monoclonal antibodies produced by cultured hybridoma on 5 and 10 percent concentration of AB , PLT and ABP compared to FCS5-10% at descending order were (1) PLT5%, (2) PLT10% , ABP5% , ABP10% , AB10%, and (3) AB5%.

Conclusions

In general FCS had the following effects on curves of cell growth: (1) the highest increase on slope of multiplication (ascending) phase, (2) the highest increase on slope of death (descending) phase, and (3) the lowest duration of stationary phase. Then, FCS can be appropriate for growth of cells at initial low cell count. Human serum AB, human platelet extract, and equal mixture of both at optimum concentrations (these supplements at high concentrations killed cells) compared to FCS showed (1) decreased slope of multiplication phase, (2) decreased slope of death phase, and (3) increased duration of stationary phase. Thus, AB and PLT may be suitable for continuous cell culture systems in which cell survival during longer times is required. Factor analysis was introduced as a model to evaluate kinetics of cell growth at different supplements.

Key words : Human serum, Cell culture, Growth kinetic, Blood groups, Factor analysis


M. Rahimkhani, Z. Alizadeh Mohammad Shir, Y. Erfani,
Volume 4, Issue 4 (2-2008)
Abstract

  Abstract

 Background and Objectives

 Bacterial contamination of blood products, especially platelets, may lead to bacterial sepsis or death and therefore is of concern. Many techniques have been explored to detect bacteria in blood products in order to prevent transfusion-related bacteria contamination and transmission. In the present study, four different methods were employed to detect 12 platelet units precontaminated with known bacteria.

 

 Materials and Methods

 Ten units of platelet concentrates were inoculated at three levels (150, 15, and 1.5 CFU per ml) with Escherichia coli and Staphyococcus epidermidis. All of the platelet concentrates and two control units of platelet concentrates were stored at 20 to 24 ° C for five days. Every morning during storage, platelet concentrates were tested for platelet pH, plasma glucose, quantitative plate culture, and gram staining on platelet centrifuged smears.

 

 Results

 Escherichia coli with 150 and 15 CFU per ml and staphylococcus epidermidis with 150 CFU per ml grew on culture medias after two days but staphylococcus epidermidis with 15 CFU per ml did after three days. The sensitivity rate of bacteria detection in platelet concentrates through gram staining was lower than quantitative culture. Despite lower plasma glucose level in platelet concentrates (as measured by hexokinase enzymatic method) inoculated with microbial staines, pH level in platelet concentrates (as measured by pH meter) contentiously increased during five days of storage.

 

 Conclusions

 The sensitivity rate of bacterial detection in platelet concentrates through measuring extra cellular pH was estimated to be higher than that of plasma glucose, culture and gram staining methods.

 

 Key words : Platelet, Glucose,Culture techniques


Dr. F. Razjou, A. Dabirmoghadam,
Volume 8, Issue 4 (1-2012)
Abstract

  Abstract

 Background and Objectives

 For reducing bacterial contamination of platelets in the medium, FDA has approved the Bact/Alert for screening the platelet units. This study attempts to compare the Bact/Alert system and the manual culture method as far as the length of time in hours of detection is concerned.

 

 Materials and Methods

 In this interventional and diagnostic study, 15 platelet units were selected randomly among 1332 units and inoculated with 10 CFU/ml of various bacteria including Streptococci, Serratia marcescens, Enterobacter cloacae, Corynebacterium diphteroid, Staphylococcus aureus and Staphylococcus epidermidis which normally contaminate platelet units. These units together with other platelet units in a blind way were tested by both Bact/Alert system and the manual method.

 

 Results

 Regarding the short expiration time of platelet units, if the length of time in hours in detection is considered as a basis for comparison, the Bact/Alert system is significantly superior to the manual method. The medium length of time in hours for detecting the aerobic bacteria by Bact/Alert system is 31 ± 8 hours, compared with the manual method which is 61 ± 11 hours. This shows that Bact/Alert system is nearly 2 times faster than the manual method.

 

 Conclusions

 Bact/Alert culture system compared with the manual method is more rapid and accurate for detection of bacterial contamination thereby improving platelet safety. Regarding serious effects of these contaminations on platelet recipients, it is also necessary to try to reduce them by using GMP.

  

  


R. Fadaei, Dr. N. Amirizadeh, Dr. M. Nikougoftar, Dr. M. Habibi Roudkenar,
Volume 9, Issue 3 (8-2012)
Abstract

  Abstract

 Background and Objectives

 The correlation between hematopoietic stem cells (HSCs) and the cells comprising the niche especially osteoblast cells is critical for maintaining stem cell activities. Yet little evidence supports the concept that HSCs regulate development of the niche. If true, it may explain why many hematopoietic defects are accompanied by changes in the osseous architecture and provide new therapeutic targets for regulating bone formation.

 

 Materials and Methods

 In this exprimental study, we cocultured bone marrow mesenchymal stem cells (MSCs) with umbilical cord blood HSCs in stem span media for 3 days. Then MSCs were differentiated to osteoblast cells by using osteogenesis kit. Evaluation of osteogenic differentiation of MSCs in different days came out to be: expression of osteopontin and osteocalsin (RT-PCR on days 4 and 6), expression of CD90 (flow cytometry on day 6), expression of alkaline phosphatase enzyme (alkaline phosphatase staining) and mineralization of MSCs (alizarin red staining on day 10).

  

 Results

 Our results showed early expression of osteopontin and osteocalsin in HSCs coculture with MSCs. These findings support the role of HSCs in induction of osteoblastic differentiation of MSCs. Decrease in the expression of CD90, higher activation in cell alkaline phosphatase enzyme, and mineralization confirmed the results.

 

 Conclusions

 In the field of human stem cells, our ex vivo findings demonstrated that HSCs can enhance differentiation of MSCs toward osteoblast cells thereby participating in formation of their niche.

  

  


Mahdi Hoseyni, Dr. E. Haddadi, Dr. M.h. Rakhshani,
Volume 15, Issue 3 (9-2018)
Abstract

Abstract
Background and Objectives
Iranian Blood Transfusion Organization as a leading and thriving organization with a scientific look at the organization's culture should enable managers to decrease the gap between the status quo and the desirable situation through identifying the employees' perceptions and expectations, prioritizing the issues, and comparing the working groups.
 
Materials and Methods
This descriptive cross-sectional study was carried out in blood centers of three provinces of Khorasan in 2015; to gather information, a 60-question questionnaire and a census sampling method were used. Further, to analyze the data, descriptive (frequency distribution, mean, standard deviation) and inferential (independent T-test, Anova, Tukey, Pearson coefficient) statistics were used based on demographic variables using SPSS and Excel softwares.
 
Results
After gathering the information of community including 78 women and 146 men, it was found that the blood centers of  the three provinces of Khorasan had a moderate and strong organizational culture with a score of 198.93.  In addition, the mission had the highest impact among different dimensions of organizational culture with the score of 50.31. However, it needs to be improved in “customer orientation” and “capacity development” indices.
 
Conclusions 
Studies show that the staff are well acquainted with the vision and strategic intentions. Hence, more trust and value should be given to the group and teamwork. In addition, with regard to the customer orientation index score, the ability of employees should be increased through holding retraining courses. And the decision makers should create an atmosphere for staff motivation, customer appreciation, and capacity development.
 

 

Dr. Davood Najafi, Dr. Ezzatollah Fathi, Dr. Raheleh Farahzadi,
Volume 15, Issue 3 (9-2018)
Abstract

Abstract
Background and Objectives
Drug resistance is one of the main challenges in the treatment of chronic myeloid leukemia (CML). In this study, with the aim of inducing apoptosis in K562 cell line (CML), this cell line was co-cultured with rat bone marrow mesenchymal stem cells (rBMSCs). Then, the rate of apoptosis as well as cell surface markers expression of CD33 and CD34 in the K562 cells line was evaluated.
 
Materials and Methods
In this experimental study, after isolating rBMSCs, the multilineag differentiation capacity of rBMSCs as well as the mesenchymal stemness of these cells was identified by evaluation of CD90, CD105, CD45, and CD56 cell surface. Then, the K562 cells were subjected for evaluation of CD33 and CD34 cell surface markers and apoptosis using a flow cytometry technique.
 
Results
Immunophenotyping of the cells indicated positive expression of CD90 and CD105 and negative expression of CD45 and CD56. The expression of CD33 and CD34 cell markers in the K562 cell line was increased 45.2% and 76.5%, respectively. Also, induction of apoptosis in the K562 cell line significantly increased (56.6%) compared to the control (p < 0.05).
 
Conclusions 
Briefly, this study showed that the co-culturing of the K562 cells line with rBMSCs causes significant increase in the expression of CD33 and CD34 cell surface markers as well as induction of apoptosis in the K562 cells (p < 0.05). This effect can be through the secreted factors and cytokines of rBMSCs; however, better evaluation in terms of type of cytokines is recommended in the future studies.
 

 

Dr. S. Milani, Dr. F. Yari,
Volume 18, Issue 2 (7-2021)
Abstract

Abstract
Background and Objectives
Mixed Lymphocytes Culture (MLC), is a method studies the cell-cell interaction between subpopulations of lymphocytes and the production of compounds caused by this interaction. MLC as a source of cytokines can increase B lymphocytes proliferation. Repeated blood transfusions cause B lymphocytes alloimmunization. Cyclosporin A has inhibitory effect on immune system. In alloimmunization, stimulated B lymphocytes can produce antibodies against exposed foreign red blood cell antigens. Immortalization of B lymphocytes can lead to continuous production of monoclonal antibodies. Considering MLC as an important source of cytokines and the presence of B lymphocytes as a group of cells in  peripheral blood cells, the aim of this study was to produce MLC and evaluate its effect on proliferation of alloimmune peripheral blood cells in the presence or absence of cyclosporine A using microscopic evaluation and dye exclusion staining method.
 
Materials and Methods
In an experimental study, RhD antigen positive and negative lymphocytes were exposed together for MLC production. Peripheral blood lymphocytes resulted from two alloimmunized patients was treated with MLC and cyclosporine, and their proliferation was analyzed compared to the control group using microscopy and trypan blue staining method. The results were analyzed by t-test using prism software (p< 0.05).
 
Results
MLC and cyclosporine increased proliferation of lymphocytes with mean value being 440000 (SD: 197989.8) vs 160000 (SD: 84852.8) of control group.
 
Conclusions 
MLC and cyclosporine increase lymphocyte cells proliferation. By confirmation of additional experiments, MLC and cyclosporine may be used as effective agents in lymphocyte proliferation and antibody production of alloimmune B lymphocytes.

Dr. M. Hoseyni, Dr. B. Hajibaygi, Dr. M. Pourmohammad, Dr. R. Mirzaie, S. Sedighzadeh, Gh. Sharifzadeh, Dr. Gh. Karimi,
Volume 21, Issue 1 (3-2024)
Abstract

Abstract
Background and Objectives
Developing a vision without understanding the organizational culture is a failed attempt. The purpose of this research is to understand organizational ­culture for understanding expectations in order to prioritize issues and compare work groups by managers for greater productivity.

Materials and Methods
This study was carried out in Iranian ­Blood­­ Transfusion Organization between 2020 and 2022. The statistical population for this research was the employees of the central headquarters and the general administrations of blood centers, and 384 of them were included in the study using stratified random sampling. To collect the required data, a 60-question organizational culture questionnaire based on Denison's model was used. For data analysis, descriptive statistics (frequency distribution, mean, standard deviation) and inferential (independent t, analysis of variance, Tukey and Pearson's coefficient) according to demographic variables, SPSS22 and Excel­ 2013 software were used.

Results
The findings show that Iranian Blood Transfusion Organization has a medium/strong organizational culture with a score of 179.31 and the highest impact after mission with a score of 46.60. Also, the indicators of "customer orientation and development of capabilities" need to be improved.

Conclusions 
According to the findings, most employees have understood the vision, strategic intentions and mission of the organization, but their capabilities are not well seen. According to the score of customer orientation, the skills and abilities of the employees should be developed by using retraining courses, while motivating the personnel, respecting the clients, and the provinces should take advantage of the experiences.


 

Dr. F. Mohammadali, Dr. S. Abroun, Dr. A. Atashi,
Volume 21, Issue 3 (10-2024)
Abstract

Abstract
Background and Objectives
Cord blood (CB) is a rich source of  Hematopoietic stem cells (HSCs). Beside the advantage the main disadvantages of CB are  the limited number of stem cells and delayed engraftment. Identifying strategies to enhance expansion and maintain stemness of HSCs can improve transplant efficiency. The goal of this study was to examine different culture conditions on HOXB4, c-Myc, Nanog, Sox2  gene expression of CB-HSCs.

Materials and Methods
In this interventional study, human cord blood CD34+ HSC, were cultured in the serum-free me­dium supplemented with cytokines (TPO, FLT3L, SCF) with/without  bone marrow mesenchymal stem cell (MSC) in 21% O2  and 5% O2 for 7 days. In day 7 the expression of, HOXB4, c-Myc, Nanog, Sox2  genes were evaluated by Real time PCR. The data analyzed using the ANOVA test  and Value < 0.05 were considered statistically significant.

Results
Highest rates of  HOXB4, c-Myc, Nanog, SOX2 mRNA were seen in coculture of HSC with bone marrow  MSC at 5% O2. Our findings demonstrated  statistically significant increase of expansion and stemness markers in 5% O2 tension versus 21%.

Conclusions 
Bone Marrow (BM) -MSC and 5% O2 combination enhanced stemness of HSC  and better mimicked the niche microenvironment conditions.

 


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