ABSTRACT
Background: The Definitive Endoderm [DE] differentiation using the undefined media and non-human feeders can cause contaminations in the generated cells for therapeutic applications. Therefore, generating safer and more appropriate DE cells is needed. This study compared five different methods to establish an appropriate method for inducing an efficient DE differentiation from Human Induced Pluripotent Stem Cells [hiPSCs] on an appropriate feeder in a more defined medium
Methods: Human Induced Pluripotent Stem Cells [hiPSCs] were cultured on inactivated feeders. Passaged hiPSCs, without feeder, were incubated for three days with Activin-A and different endodermal differentiation media including 1-FBS, 2-B27, 3- ITS and albumin fraction-V, 4-B27 and ITS and 5-like the third medium. The feeder cells in the first four methods were Mouse Embryonic Fibroblasts [MEFs] and in the fifth method were human adult bone marrow Mesenchymal Stem Cells [hMSCs]. DE markers FOXA2, SOX17 and CXCR4 and also pluripotency marker OCT4 were evaluated using qRT-PCR, as well as FOXA2 by the immunocytochemistry
Results: QRT-PCR analysis showed that after three days, the expression levels of DE and pluripotency markers in the differentiated hiPSCs among all five groups did not have any significant differences. Similarly, the immunocytochemistry analysis demonstrated that the differentiated hiPSCs expressed FOXA2, with no significant differences
Conclusion: Despite this similarity in the results, the third differentiation medium has more defined and cost effective components. Furthermore, hMSC, a human feeder, is safer than MEF. Therefore, the fifth method is preferable among other DE differentiation methods and can serve as a fundamental method helping the development of regenerative medicine
ABSTRACT
Background: About 8 percent of all cancers in human population are related to leukemia and it is one of the most common malignancies in children. The aim of this study was to compare the prevalence of age, gender and blood group types with the frequency of leukemia among the children with leukemia in Qazvin province during the 2006 to 2016
Materials and Methods: This was a cross-sectional analysis. Investigated population was 110 children and adolescents under 18 years in the hospitals of Qazvin province. The date collecting method was through review of medical records of the patients and their analysis performed by using SPSS version 16
Results: According to data from this study, leukemia ALL-L1 is more frequent in Qazvin than other types of leukemia, and children with ages 0-5 years was more than other age groups. This disorder is more common in boys than girls, and among the patients, the people who has A and O blood groups, and Rh + are the most abundant
Conclusion: such factors like age, gender and blood groups can use as prognostic factors in children leukemia. So that leukemia in children less than 5 years old is more than any other age. In addition to that; the incidence of leukemia ALL-L1 reduced with increasing age in the general population in Qazvin and number of boys with leukemia is more than girls
ABSTRACT
Several influential factors such as transcription factors and intracellular signaling components are involved in differentiation of stem cells into a specific lineage. P15 and p16 proteins are among these factors. Accumulating evidences has introduced the epigenetic as a master regulator of these factors during lineage specification. The main objective of this study is to determine the correlation between the expression level and methylation pattern of P15 and P16 genes in erythroid lineage after in vitro differentiation by erythropoietin [EPO]. The purified and expanded CD34+ cord blood stem cells were differentiated into erythroid lineage in the presence of EPO. DNA was isolated from both cord blood stem cells and differentiated cells. The Real-Time PCR performed using cDNA and the isolated DNA was used in methylation Specific PCR [MSP] reaction for methylation pattern analysis in both pre and post differentiation stages. The study demonstrated that P15 and P16 genes have partial methylation after erythroid differentiation by EPO. The Expression of P15 gene was higher after differentiation and the expression of P16 gene had a slightly decreased level in post differentiation stage. Significant increase in P15 gene expression after differentiation to erythroid lineage, suggests the remarkable efficacy of this gene in erythroid function. According to upregulation of P15 gene after differentiation despite unchanged methylation status and slight down regulation of P16 gene with slight hyper-methylation of the gene it can be suggested that although the methylation can affects the expression level of P16 gene, the P15 gene is not affected by this mechanism during erythroid differentiation mediated by EPO