The Production and Evaluation of an Electrochemical Sensors for Strychnine and Its Main Metabolite Strychnine N-Oxide for Their Use in Biological Samples.

Strychnine (STN) and its major metabolite Strychnine N-Oxide (SNO) were examined electrochemically. Both parent compounds and its major metabolite showed electroactivity on glassy carbon electrodes using CV and DPV techniques. One oxidation peak at 1008 mV was observed for STN with the optimum peak intensity at pH 7. SNO produced two oxidation peaks, at 617 mV and 797 mV, at pH 5. The peaks demonstrated irreversible behaviour and the irreversibility of the system was confirmed at different scan rates. A calibration curve was produced for both CV and DPV measurements and the sensitivity of the proposed EC method was good compared with previous electrochemical and non-electrochemical methods. The precision of oxidation peak of STN using the STN-MIP method produced a maximum value of 11.5% and 2.32% for inter-day and intraday %RSD, respectively. The average% recovery was around 92%. The electrochemical method has been successfully applied to the determination of STN in spiked plasma and urine samples. For SNO, both anodic peaks of SNO demonstrated irreversible behaviour. A different sweep rate was used for calculating the number of 'transfer electrons' in the system; based on this, the mechanism of oxidation reaction was proposed. Calibration curves for both oxidative peaks were produced using DPV measurements. The second anodic peak demonstrated high linearity and precision with %RSD < 1.96%.

Computational Design of a Molecularly Imprinted Polymer for the Biomonitoring of the Organophosphorous Metabolite Chlorferron.

Coumaphos is an organophosphorus compound used as insecticide and frequently used by beekeepers for the management of parasitic mites. The most important metabolite, chlorferron (CFN), has been identified in biological samples and foodstuff. The need to quickly identify the presence of typical metabolites, as an indication of interaction with coumaphos has driven the need to produce a highly sensitive electrochemical method for chlorferron analysis, based on molecularly imprinting polymers (MIP) technology. It showed irreversible behaviour with mixed diffusion/adsorption-controlled reactions at the electrode surface. A monoelectronic mechanism of reaction for oxidation has also been suggested. The linear range observed was from 0.158 to 75 µM. Median precision in terms of %RSD around 3% was also observed. For DPV, the limit of detection (LOD) and the limit of quantitation (LOQ) for the CFN-MIP were 0.158 µM and 0.48 µM, respectively. The obtained median % recovery was around 98%. The results were also validated to reference values obtained using GC-MS. Urine and human synthetic plasma spiked with CFN were used to demonstrate the usability of the method in biological samples, showing the potential for biomonitoring. The developed imprinted sensor showed maximum signal change less than 16.8% when related metabolites or pesticide were added to the mix, suggesting high selectivity of the MIP sensor toward CFN molecules. The results from in vitro metabolism of CMP analysed also demonstrates the potential for detection and quantification of CFN in environmental samples. The newly developed CFN-MIP sensor offers similar LoDs than chromatographic methods with shorter analysis time.

Anti-angiogenic drugs: direct anti-cancer agents with mitochondrial mechanisms of action.

Components of the mitochondrial electron transport chain have recently gained much interest as potential therapeutic targets. Since mitochondria are essential for the supply of energy that is required for both angiogenic and tumourigenic activity, targeting the mitochondria represents a promising potential therapeutic approach for treating cancer. Here we investigate the established anti-angiogenesis drugs combretastatin A4, thalidomide, OGT 2115 and tranilast that we hypothesise are able to exert a direct anti-cancer effect in the absence of vasculature by targeting the mitochondria. Drug cytotoxicity was measured using the MTT assay. Mitochondrial function was measured in intact isolated mitochondria using polarography, fluorimetry and enzymatic assays to measure mitochondrial oxygen consumption, membrane potential and complex I-IV activities respectively. Combretastatin A4, OGT 2115 and tranilast were both shown to decrease mitochondrial oxygen consumption. OGT 2115 and tranilast decreased mitochondrial membrane potential and reduced complex I activity while combretastatin A4 and thalidomide did not. OGT 2115 inhibited mitochondrial complex II-III activity while combretastatin A4, thalidomide and tranilast did not. Combretastatin A4, thalidomide and OGT 2115 induced bi-phasic concentration-dependent increases and decreases in mitochondrial complex IV activity while tranilast had no evident effect. These data demonstrate that combretastatin A4, thalidomide, OGT 2115 and tranilast are all mitochondrial modulators. OGT 2115 and tranilast are both mitochondrial inhibitors capable of eliciting concentration-dependent reductions in cell viability by decreasing mitochondrial membrane potential and oxygen consumption.

The role of Nanog expression in tamoxifen-resistant breast cancer cells.

There is an accumulation of evidence that shows a significant role of cancer stem cells in tumor initiation, proliferation, relapse, and metastasis. Nanog is the most important core transcription marker of stem cells, known by its role in maintaining pluripotency, proliferation, and differentiation. Therefore, this study aimed to examine the role of Nanog in breast cancer cell tamoxifen resistance and its implications in breast cancer treatment. In this study, the expression of the three core transcription markers Nanog, Oct3/4, and Sox2 were quantitatively evaluated using flow cytometry. Then, small interfering RNA (siRNA) against human Nanog was transfected into tamoxifen-resistant breast cancer cells via Lipofectamine 2000. Nanog gene expression in the cells was detected using reverse transcription polymerase chain reaction (RT-PCR). The change in cell proliferation was evaluated using the tetrazolium bromide method. An enzyme-linked immunosorbent assay was used to detect apoptosis of the transfected cells alone and in combination with 4-hydroxytamoxifen. The results showed a high level expression of Nanog, Oct3/4, and Sox2 in MDA-MB-231 and MCF7/tamoxifen resistant cells compared with MCF7/wild-type. siRNA-mediated Nanog gene silencing can efficiently inhibit cell proliferation and induce apoptosis of tamoxifen-resistant breast cancer cells. This study provides a basis for further study of the role of Nanog in developing resistance to tamoxifen, its implication in breast cancer management, and as a new strategy to enhance response to endocrine therapy.

The power of the age standardized incidence rate to discover the gene link between cancer diseases: development of a new epidemiological method to save money, time, and effort for genetic scientists.

BACKGROUND: This study provides an incipient epidemiological rule using the concept of direct method of standardization to determine the genetic link between cancer diseases. METHODS: The overall 8 or 10 years age standardized incidence rate (ASIR) for both cancer diseases, for example (A) and (B) should be calculated for all regions of the country. A line chart should be used to display the overall ASIR trend of both diseases (A and B). Pearson's correlation can be used to determine the strength of the association between the overall ASIRs of both diseases. The overlap or opposite direction of the overall ASIR trend of both diseases (A and B) should be determined and studied for possible associations between cancer diseases. RESULTS: If the trend of the overall 8 or 10 years ASIR of a disease (A) follows that of disease (B) in all regions of the country, then the genes of patients with both diseases (A and B) will be highly homogeneous, and they should be studied in the region with the highest and lowest overall ASIR for both diseases (A and B). In addition, if there is an opposite direction or overlapping trend for both diseases (A and B) in certain regions of the country or among specific groups of people with the same demographic characteristics, then the genes of patients will be investigated for both diseases to identify the potential gene link between cancer diseases. CONCLUSION: This study revealed that the overall ASIR trends of female breast cancer, prostate cancer, and ovarian cancer are very similar in all regions of Saudi Arabia and England. Our epidemiological evidence helps to save money, time, and effort for testing the potential gene link between cancer diseases.

The incidence rate of thyroid cancer among women in Saudi Arabia: an observational descriptive epidemiological analysis of data from Saudi Cancer Registry 2001-2008.

This study provides a descriptive epidemiological data of thyroid cancer cases diagnosed from 2001 to 2008 among Saudi women, including the frequency and percentage of cases, the crude incidence rate (CIR) and the age-standardised incidence rate (ASIR) stratified by the region and year of diagnosis. This is a retrospective descriptive epidemiological analysis of all Saudi thyroid cancer cases recorded in the Saudi Cancer Registry (SCR) between January 2001 and December 2008. The statistical analyses were applied using descriptive statistics with the Statistical Package for the Social Sciences version 20.0. A total of 2,930 cases were recorded in the SCR between January 2001 and December 2008. The region of Riyadh in Saudi Arabia had the highest overall ASIR at 9.43 per 100,000 women, followed by Tabuk at 7.11 and eastern region at 6.5, while Jazan and Jouf had the lowest average ASIRs at 1.97 and at 2.72, respectively. The region of Qassim recorded the greatest changes of ASIR at 5.5 per 100,000 women from 2001 to 2008. There was a slight increase in the CIRs and ASIRs for thyroid cancer in Saudi Arabia between 2001 and 2008. Riyadh, Tabuk and eastern region were the highest overall ASIR in Saudi Arabia. While, Jazan and Hail had the lowest rates. Finally, the region of Qassim had the highest changes in CIR and ASIR from 2001 to 2008. Further analytical studies are needed to determine the potential risk factors of thyroid cancer disease among Saudi women.

The pattern of Middle East respiratory syndrome coronavirus in Saudi Arabia: a descriptive epidemiological analysis of data from the Saudi Ministry of Health.

PURPOSE: This study describes the epidemiology of Middle East respiratory syndrome coronavirus (MERS-CoV) in Saudi Arabia. PATIENTS AND METHODS: Epidemiological analysis was performed on data from all MERS-CoV cases recorded by the Saudi Ministry of Health between June 6, 2013 and May 14, 2014. The frequency of cases and deaths was calculated and adjusted by month, sex, age group, and region. The average monthly temperature and humidity of infected regions throughout the year was also calculated. RESULTS: A total of 425 cases were recorded over the study period. The highest number of cases and deaths occurred between April and May 2014. Disease occurrence among men (260 cases [62%]) was higher than in women (162 cases [38%]), and the case fatality rate was higher for men (52%) than for women (23%). In addition, those in the 45-59 years and ≥60 years age groups were most likely to be infected, and the case fatality rate for these people was higher than for other groups. The highest number of cases and deaths were reported in Riyadh (169 cases; 43 deaths), followed by Jeddah (156 cases; 36 deaths) and the Eastern Region (24 cases; 22 deaths). The highest case fatality rate was in the Eastern Region (92%), followed by Medinah (36%) and Najran (33%). MERS-CoV infection actively causes disease in environments with low relative humidity (<20%) and high temperature (15°C-35°C). CONCLUSION: MERS-CoV is considered an epidemic in Saudi Arabia. The frequency of cases and deaths is higher among men than women, and those above 45 years of age are most affected. Low relative humidity and high temperature can enhance the spread of this disease in the entire population. Further analytical studies are required to determine the source and mode of infection in Saudi Arabia.

The incidence of leukemia in Saudi Arabia. Descriptive epidemiological analysis of data from the Saudi Cancer Registry 2001-2008.

OBJECTIVE: To describe the epidemiological data of leukemia cases diagnosed from 2001 to 2008 among male and female Saudis, including the frequency and percentage of cases, the crude incidence rate (CIR), and the age-standardized incidence rate (ASIR) stratified by leukemia subtype, region, and year of diagnosis. METHODS: This is a retrospective descriptive epidemiological analysis of all Saudi leukemia cases recorded in the Saudi Cancer Registry SCR between January 2001 and December 2008. The study was carried out in 2013 to investigate the pattern of leukemia in the Saudi population. Descriptive statistics and Poisson regression model were used. RESULTS: A total of 3852 leukemia cases were registered in the SCR between January 2001 and December 2008. The region of Riyadh, Saudi Arabia had the highest overall ASIR among Saudi males at 5.2 per 100,000 males, followed by both the Eastern region and Northern region at 4.9 per 100,000 males. Furthermore, the region of Najran recorded the highest overall ASIR among Saudi females at 4.5 per 100,000 females. However, Jazan had the lowest average ASIRs of leukemia in Saudi Arabia. CONCLUSION: There was a slight increase in the CIRs and ASIRs of leukemia in Saudi Arabia between 2001 and 2008. Riyadh, the Eastern region, and the Northern region had the highest overall ASIRs of leukemia among Saudi males, and Najran had the highest overall ASIRs of leukemia among Saudi females; while Jazan had the lowest rates among the Saudi population.

Incidence rate of non-Hodgkin's lymphomas among males in Saudi Arabia: an observational descriptive epidemiological analysis of data from the Saudi Cancer Registry, 2001-2008.

BACKGROUND: This study describes epidemiological data of non-Hodgkin's lymphoma (NHL) diagnosed from 2001 to 2008 among Saudi men. MATERIALS AND METHODS: Retrospective data from all NHL cancer cases among Saudi men recorded in the Saudi Cancer Registry (SCR) between January 2001 and December 2008 were used. Descriptive statistics, analysis of variance, Poisson regression, and simple linear regression were also used. RESULTS: In total, 2,555 new cases of NHL were recorded between January 2001 and December 2008. The region of Riyadh, Saudi Arabia had the highest overall age-standardized incidence rate (ASIR) at 7.8, followed by the Eastern region at 6.8, and Makkah at 6.1 per 100,000 men; however, Jazan, Hail, and Baha had the lowest average ASIRs at 2.5, 3.7, and 3.9 per 100,000 men, respectively. The incidence-rate ratio for the number of NHL cases was significantly higher in Riyadh (4.68, 95% confidence interval [CI] 4.11-5.32), followed by Makkah (4.47, 95% CI 3.94-5.07), and the Eastern region of Saudi Arabia (3.27, 95% CI 2.90-3.69) than that in the reference region of Jazan. Jouf had the highest changes in the ASIRs of NHL among Saudi men from 2001 and 2008 (5.0 per 100,000 men). CONCLUSION: A significant increase in the crude incidence rate and ASIR for NHL in Saudi Arabia between 2001 and 2008 was found. Riyadh, the Eastern region, and Makkah had the highest overall ASIR in Saudi Arabia. Jazan, Hail, and Baha had the lowest rates. Additionally, Riyadh, Makkah, and the Eastern region had the highest incidence-rate ratio for the number of NHL cases. Finally, Jouf had the highest changes in crude incidence rate and ASIR from 2001 to 2008. Further analytical studies are needed to determine the potential risk factors of NHL among Saudi men.

Incidence rate of ovarian cancer cases in Saudi Arabia: an observational descriptive epidemiological analysis of data from Saudi Cancer Registry 2001-2008.

PURPOSE: This study provides descriptive epidemiological data, such as the percentage of cases diagnosed, crude incidence rate (CIR), and age-standardized incidence rate (ASIR) of ovarian cancer in Saudi Arabia from 2001-2008. PATIENTS AND METHODS: A retrospective descriptive epidemiological analysis of all ovarian cancer cases recorded in the Saudi Cancer Registry (SCR) from January 2001-December 2008 was performed. The data were analyzed using descriptive statistics, analysis of variance tests, Poisson regression, and simple linear modeling. RESULTS: A total of 991 ovarian cancer cases were recorded in the SCR from January 2001-December 2008. The region of Riyadh had the highest overall ASIR at 3.3 cases per 100,000 women, followed by the Jouf and Asir regions at 3.13 and 2.96 cases per 100,000 women. However, Hail and Jazan had the lowest rates at 1.4 and 0.6 cases per 100,000 women, respectively. Compared to Jazan, the incidence rate ratio for the number of ovarian cancer cases was significantly higher (P<0.001) in the Makkah region at 6.4 (95% confidence interval [CI]: 4.13-9.83), followed by Riyadh at 6.3 (95% CI: 4.10-9.82), and the eastern region of Saudi Arabia at 4.52 (95% CI: 2.93-6.98). The predicted annual CIR and ASIR for ovarian cancer in Saudi Arabia could be defined by the equations 0.9 + (0.07× years) and 1.71 + (0.09× years), respectively. CONCLUSION: We observed a slight increase in the CIRs and ASIRs for ovarian cancer in Saudi Arabia from 2001-2008. Riyadh, Jouf, and Asir had the highest overall ASIR, while Jazan and Hail had the lowest rates. Makkah, Riyadh, and the eastern region of Saudi Arabia had the highest incidence rate ratio for the number of ovarian cancer cases. Further analytical studies are required to determine the potential risk factors of ovarian cancer among Saudi women.

The incidence rate of corpus uteri cancer among females in Saudi Arabia: an observational descriptive epidemiological analysis of data from Saudi Cancer Registry 2001-2008.

BACKGROUND: The present study reviews the epidemiological data on corpus uteri cancer among Saudi women, including its frequency, crude incidence rate, and age-standardized incidence rate (ASIR), adjusted by region and year of diagnosis. METHODS: A retrospective, descriptive epidemiological analysis was conducted of all the corpus uteri cancer cases recorded in the Saudi Cancer Registry between January 2001 and December 2008. The statistical analyses were performed using descriptive statistics, analysis of variance, Poisson regression, and a simple linear model. RESULTS: A total of 1,060 corpus uteri cancer cases were included. Women aged 60-74 years of age were most affected by the disease. The region of Riyadh in Saudi Arabia had the highest overall ASIR, at 4.4 cases per 100,000 female patients, followed by the eastern region, at 4.2, and Makkah, at 3.7. Jazan, Najran, and Qassim had the lowest average ASIRs, ranging from 0.8 to 1.4. A Poisson regression model using Jazan as the reference revealed that the corpus uteri cancer incidence rate ratio was significantly higher for the regions of Makkah, at 16.5 times (95% confidence interval [CI]: 8.0-23.0), followed by Riyadh, at 16.0 times (95% CI: 9.0-22.0), and the eastern region, at 9.9 times (95% CI: 5.6-17.6). The northern region experienced the highest changes in ASIRs of corpus uteri cancer among female Saudi patients between 2001 and 2008. CONCLUSION: There was a slight increase in the crude incidence rates and ASIRs for corpus uteri cancer in Saudi Arabia between 2001 and 2008. Older Saudi women were most affected by the disease. Riyadh, the eastern region, and Makkah had the highest overall disease ASIRs and incidence rate ratios, while Jazan, Najran, and Qassim had the lowest rates. Finally, the northern region experienced the greatest changes in ASIR during the studied period. Further analytical studies are necessary to determine potential risk factors of corpus uteri cancer among female Saudi patients.

The incidence rate of prostate cancer in Saudi Arabia: an observational descriptive epidemiological analysis of data from the Saudi Cancer Registry 2001-2008.

BACKGROUND: This study presents descriptive epidemiological data related to prostate cancer cases diagnosed from 2001 to 2008 among Saudi men, including the frequency and percentage of cases, the crude incidence rate (CIR) and the age-standardised incidence rate (ASIR) adjusted by region and year of diagnosis. METHODS: This is a retrospective descriptive epidemiological analysis of all Saudi prostate cancer cases recorded in the Saudi Cancer Registry (SCR) between January 2001 and December 2008. Statistical analyses were conducted using descriptive statistics, the Poisson regression model, a simple linear regression model and analysis of variance with the Statistical Package for the Social Sciences version 20.0 (SPSS). RESULTS: A total of 1739 cases were registered in the SCR between January 2001 and December 2008. The eastern region of Saudi Arabia had the highest overall ASIR at 10.1 per 100,000 men, followed by Riyadh at 7.1 and Makkah at 5.2, while Jizan and Najran had the lowest average ASIRs at 1.4 and at 2.0, respectively. The Poisson regression model for the region of Jizan shows that the incidence rate ratio (IRR) was significantly higher (p < 0.001); for the regions of Riyadh at 3.43 times (95% CI, 3.13-3.73), followed by Makkah at 2.56 (95% CI, 2.38-2.74) and the eastern province at 1.93 (95% CI, 1.64-2.23). The annual prediction of the CIR of prostate cancer in Saudi Arabia could be defined by the equation 1.53 + (0.094 × Years), and the prediction of the ASIR could be defined by the equation 2.98 + (0.182 × Years). CONCLUSION: There was a steady increase in the CIRs and ASIRs for prostate cancer between 2001 and 2008. The eastern region, Riyadh and Makkah had the highest overall ASIR in Saudi Arabia. Further effective screening programmes and active treatment measures are needed to control prostate cancer disease among Saudi men.

The incidence rate of female breast cancer in Saudi Arabia: an observational descriptive epidemiological analysis of data from Saudi Cancer Registry 2001-2008.

BACKGROUND: This study presents descriptive epidemiological data related to breast cancer cases diagnosed from 2001 to 2008 among Saudi women, including the frequency and percentage of cases, the crude incidence rate (CIR), and the age-standardized incidence rate (ASIR), adjusted by the region and year of diagnosis. METHODS: This is a retrospective descriptive epidemiological study of all Saudi female breast cancer cases from 2001 to 2008. The statistical analyses were conducted using descriptive statistics, a linear regression model, and analysis of variance with the Statistical Package for the Social Sciences version 20.0. RESULTS: A total of 6,922 female breast cancer cases were recorded in the Saudi Cancer Registry from 2001 to 2008. The highest overall percentages (38.6% and 31.2%) of female breast cancer cases were documented in women who were 30-44 and 45-59 years of age, respectively. The eastern region of Saudi Arabia had the highest overall ASIR, at 26.6 per 100,000 women, followed by Riyadh at 20.5 and Makkah at 19.4. Jazan, Baha, and Asir had the lowest average ASIRs, at 4.8, 6.1, and 7.3 per 100,000 women, respectively. The region of Jouf (24.2%; CIR 11.2, ASIR 17.2) had the highest changes in CIR and ASIR from 2001 to 2008. While Qassim, Jazan, and Tabuk recorded down-trending rates with negative values. CONCLUSION: There was a significant increase in the CIRs and ASIRs for female breast cancer between 2001 and 2008. The majority of breast cancer cases occurred among younger women. The region of Jouf had the greatest significant differences of CIR and ASIR during 2001 to 2008. Jazan, Baha, and Najran had the lowest average CIRs and ASIRs of female breast cancer, whereas the linear trend upward is a concern in certain regions, such as the eastern region, Makkah, and Riyadh. However, further analytical epidemiological research is needed to identify the potential risk factors involved in the increase in the prevalence of breast cancer among Saudi women.

The stem cell factor antibody enhances the chemotherapeutic effect of adriamycin on chemoresistant breast cancer cells.

BACKGROUND: The outcome of chemotherapy in breast cancer is strongly influenced by multidrug resistance (MDR). Several surrogate markers of chemoresistance have been identified including - CD24 (cluster differentiation 24) expression, stem cell growth factor (SCF), B-cell lymphocyte protein 2 (Bcl-2) and annexin V. The present study aimed to examine the expression of CD24 in the sensitive breast cancer cell line MCF-7 (Michigan Foudation-7) and MCF-7/adriamycin resistant (MCF-7/AdrRes) cells, and, if minimal effective doses of the anthracycline drug adriamycin (0.579 µM and 88.2 µM) would be enhanced by the antibody to SCF (anti-SCF). METHODS: CD24 expression was analysed by flow cytometry. Both Bcl-2 and annexin V protein expression were quantitatively assessed by the enzyme-linked immunosorbent assay (ELISA). RESULTS: In MCF-7/AdrRes cells the expression of CD24 was significantly higher compared to MCF-7 cells, 86.6% and 16.3% (p < 0.001), respectively. Bcl-2 expression was significantly increased in the presence of adriamycin and SCF (p < 0.038) and decreased in the presence of adriamycin and anti-SCF. When adriamycin, anti-SCF and SCF were combined or when adriamycin was used alone the decrease in Bcl-2 expression was insignificantly altered. In the presence of both adriamycin and SCF the expression of annexin V was decreased. However, it was significantly increased in the presence of adriamycin and anti-SCF (p < 0.042), as well as adriamycin, anti-SCF and SCF combined.In MCF-7 cells the effect of adriamycin alone or with either SCF, anti-SCF or anti-SCF or SCF combined, did not significantly alter the expression of Bcl-2. However, in the presence of both adriamycin and SCF the expression of annexin V was decreased, but was significantly increased in the presence of adriamycin and anti-SCF (p < 0.001), adriamycin, anti-SCF and SCF combined and adriamycin alone. Our results demonstrate that anti-SCF with low dose of adriamycin reduces Bcl-2 expression in MCF-7/AdrRes cells and increases annexin V expression in both MCF7/AdrRes and MCF-7 cells. CONCLUSION: Adding anti-SCF to the chemotherapeutic regime of adriamycin may strongly enhance its chemotherapeutic effect in the treatment of patients with breast cancer.

New approach to isolate mesenchymal stem cell (MSC) from human umbilical cord blood.

HUCB (human umbilical cord blood) has been frequently used in clinical allogeneic HSC (haemopoietic stem cell) transplant. However, HUCB is poorly recognized as a rich source of MSC (mesenchymal stem cell). The aim of this study has been to establish a new method for isolating large number of MSC from HUCB to recognize it as a good source of MSC. HUCB samples were collected from women following their elective caesarean section. The new method (Clot Spot method) was carried out by explanting HUCB samples in mesencult complete medium and maintained in 37°C, in a 5% CO2 and air incubator. MSC presence was established by quantitative and qualitative immunophenotyping of cells and using FITC attached to MSC phenotypic markers (CD29, CD73, CD44 and CD105). Haematopoietic antibodies (CD34 and CD45) were used as negative control. MSC differentiation was examined in neurogenic and adipogenic media. Immunocytochemistry was carried out for the embryonic markers: SOX2 (sex determining region Y-box 2), OLIG-4 (oligodendrocyte-4) and FABP-4 (fatty acid binding protein-4). The new method was compared with the conventional Rosset Sep method. MSC cultures using the Clot Spot method showed 3-fold increase in proliferation rate compared with conventional method. Also, the cells showed high expression of MSC markers CD29, CD73, CD44 and CD105, but lacked the expression of specific HSC markers (CD34 and CD45). The isolated MSC showed some differentiation by expressing the neurogenic (SOX2 and Olig4) and adipogenic (FABP-4) markers respectively. In conclusion, HUCB is a good source of MSC using this new technique.