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Showing 3 results for Colony
Synthesis of human placental CDNA and demonstration of the expression of M-CSF in that tissue
R Arvan , E Elahy ,
Volume 56, Issue 5 (7-1998)
Abstract
Macrophage colony stimulating factor (M-CSF) has previously been shown to affect the differentiation of cells of the mono-nuclear phagocytic line. More recent studies indicate that M-CSF may have a role in pregnancy. In the present study, the expression of M-CSF in the human placenta was demonstrated. Placental mRNA was isolated and used as template for synthesis of complementary DNA (cDNA). The presence of M-CSF related sequences in the cDNA was shown by PCR and RT-PCR reactions in which M-CSF specific primers were used. In addition, it was shown that a 2.4 kb cDNA after electrophoresis and transfer to a nylon filter, hybridized with a digoxygenin labeled M-CSF specific probe.
In-vitro differentiation of human embryonic stem cells into hemangioblasts
Ganji Fatemeh, Abruon Saeid, Baharvand Hossein, Ebrahimi Marzieh, Aghdami Nasser,
Volume 70, Issue 3 (6-2012)
Abstract

Background: Human embryonic stem cells (hESCs) are capable of self-renewal and large-scale expansion. They also have the capacity to differentiate into a variety of cell types including liver, cardiac and neuron cells. However, it is not yet clear whether hESCs can differentiate to hemangioblasts under in-vitro conditions. Hemangioblasts are bipotential progenitors that can generate hematopoietic lineages and endothelial cells. The aim of this study was to identify the potential of human Royan H5 embryonic stem cells in differentiating into hemangioblast cells.

Methods: HESCs were cultured at suspension system in DMEM/F12 supplemented with bFGF. 7-day old cells differentiated into blast cells under defined condition consisting of hematopoietic cytokines including BMP4, VEGF, etc. Blast cell markers kinase insert domain receptor (KDR), CD31, and CD34 were evaluated by flow cytometry and blast gene expressions (TAL-1, Runx-1 and CD34) were detected by qRT-PCR. Clonogenic assays were performed in semisolid medium by colony forming unit-assays.

Results: The hESCs (Royan H5) had the capacity of differentiating into hemangioblast cells. We could detect colonies that expressed 79%±12.5 KDR+, 5.6%±2.8 CD31+-CD34+ and 6%±2.12 KDR+-CD31+ on day 8 in the hESCs. Up-regulation of TAL-1, Runx-1 and CD34 occurred during hemangioblast commitment (P≤0.05 and P≤0.01, respectively). Moreover, hemangioblast cells generated mixed-type and endothelial-like colonies in semi-solid media.

Conclusion: Our results showed that hESCs (Royan H5) were able to differentiate into hemangioblasts under in-vitro conditions. The hemangioblasts had the potential to generate two non-adherent (Mixed-type) and adherent (endothelial-like) cell populations.


Comparison of MTT, trypan blue, and clonogenic assay, to determine the viability in human anaplastic thyroid cancer cell line
Marjan Ghorbani-Anarkooli , Sara Dabirian, Hasan Moladoust, Adib Zendedel, Mohammad Hadi Bahadori,
Volume 77, Issue 1 (4-2019)
Abstract
Background: Evaluation of cell viability is momentous in pharmacologic and oncological research. Cell viability evaluation determines cell sensitivity and consequently treatment outcome. Various methods are available to determine cell survival. Each of these methods evaluates different endpoints. Accordingly, determining the correlation between these methods is important. In this study, in order to determine the viability of human anaplastic thyroid cancer cell line, the sensitivity of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, trypan blue test and clonogenic assay were compared.
Methods: This experimental study was performed in the Cellular and Molecular Research Center at Guilan University of Medical Sciences, Rasht, Iran from October 2016 to March 2017. The human anaplastic thyroid cancer cell line was cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS). The cultured cells were treated with melatonin, for 24 hours. Then, the viability of the cells was evaluated by MTT assay, trypan blue test and clonogenic assay. Furthermore, plating efficiency and surviving fraction were used in order to draw survival curve in the clonogenic assay.
Results: The concentration of melatonin at IC50 point was 4.794±0.117 millimolar (mM) in MTT assay, 4.375±0.894 mM in trypan blue test and 2.246±0.326 mM in clonogenic assay. Comparing the IC50 values of these test revealed that C50 values obtained from MTT assay and trypan blue test had no significant difference (P=0.6446), while there was a significant difference between IC50 values obtained from MTT and clonogenic assays (P=0.0032). Moreover, the IC50 values obtained from trypan blue test and clonogenic assay were also significantly different (P=0.0078). The results of the regression analysis of cell viability were shown a linear, positive and significant correlation between these three methods and MTT assay and trypan blue test showed higher correlation (r=0.99, P<0.001).
Conclusion: Based on our results, all these methods were effective to identify cytotoxicity in human anaplastic thyroid cancer cell line, while MTT assay and trypan blue test were more sensitive than clonogenic assay.

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