The in vitro treatment of mesenchymal stem cells for colorectal cancer cells.

Colorectal cancer is the most common tumor of the gastrointestinal system. The conventional treatment options for colorectal cancer are troublesome for both patients and clinicians. Recently, mesenchymal stem cells (MSCs) have been the novel focus for cell therapy due to their migration to tumor sites. In this study, the apoptotic effect of MSCs on colorectal cancer cell lines has been aimed. HCT-116 and HT-29 were selected as the colorectal cancer cell lines. Human umbilical cord blood and Wharton's jelly were used as mesenchymal stem cell sources. To discriminate against the apoptotic effect of MSC on cancer, we also used peripheral blood mononuclear cells (PBMC) as a healthy control group. Cord blood-MSC and PBMC were obtained by ficoll-paque density gradient, and Wharton's jelly-MSC by explant method. Transwell co-culture systems were used as cancer cells or PBMC/MSCs at ratios of 1/5 and 1/10, with incubation times of 24 h and 72 h. The Annexin V/PI-FITC-based apoptosis assay was performed by flow cytometry. Caspase-3 and HTRA2/Omi proteins were measured by ELISA. For both ratios in both cancer cells, it was found that the apoptotic effect of Wharton's jelly-MSC was significantly higher in 72-h incubations (p < 0.006), whereas the effect of cord blood mesenchymal stem cell in 24-h incubations were higher (p < 0.007). In this study, we showed that human cord blood and tissue-derived MSCs treatment led to colorectal cancers to apoptosis. We anticipate that further in vivo studies may shed light on the apoptotic effect of MSC.

Increasing Apoptotic Effect of Cord Blood and Wharton's Jelly-derived Mesenchymal Stem Cells on HT-29.

BACKGROUND: Colorectal cancer (CRC) is the third most common cancer worldwide. Recently, mesenchymal stem cells (MSCs) have been considered a suitable cell therapy option for cancer due to their high migration rate to the tumor site. OBJECTIVES: The study aimed to compare the effects of human umbilical cord blood derived-MSC (UCMSC) and human Wharton's Jelly derived-MSC (WJ-MSC) on the HT-29 cell line. METHODS: UC-MSC was obtained by Ficoll-Paque density gradient and WJ-MSC by explant method. The characterizations of MSCs and apoptosis assays were performed by flow cytometry, and caspase-3 protein levels were measured by ELISA. RESULTS: After 72 hours of HT-29 cancer cells incubation, it was indicated that WJ-MSC was more effective at 1:5 and 1:10 ratios. Similar results were found for caspase-3 by ELISA. Moreover, WJ-MSC (1:5, p < 0.006; 1:10, p < 0.007) was found to be more effective at both doses compared to UC-MSC. CONCLUSION: In this study, we used two different MSC sources at two different ratios to evaluate the apoptotic effect of MSC in vitro on HT-29 CRC cells. As a result, WJ-MSC indicated a more apoptotic effect on HT-29 cells compared to CB-MSC. We anticipated that this preliminary in vitro study would be extended in future in vitro/in vivo studies. Moreover, investigating the behavior of MSC in colorectal tumor microenvironment will be beneficial for the stem cell therapy approach.

Blood and bone marrow donor registry of Istanbul medical faculty activity and experience in past 3 years : A cross-sectional documentation of TRIS activity: the first blood and marrow registry in Turkey.

Allogeneic stem cell transplantation (SCT) offers a potential cure for some hematological malignancies. For those patients without a family donor, unrelated donor (MUD) registries serve for identifying the best donor. In the present study, we aimed to give a cross-sectional report of our registry's activity and experience as the first established national MUD registry in the country. The study is retrospective and covers the period of 2016 to 2019. A total of 1855 donor searches were performed, and 642 were included in the study. All data were electronically obtained from the institutional database system. All SCTs were either 10/10 or 9/10 HLA matched and originated from an international registry. The most preferred stem cell source was peripheral blood (70.2%). A quarter of transplants were performed using bone marrow, and cord blood was used with a rate of 1.4%. The pandemic-related problems were similar for the other two national registries. During the pandemic, 71 of 432 patients who were searched for donors underwent stem cell transplant(SCT). The low number was related mostly with postponing of SCTs and/also difficulties in continuing of volunteering and in achievement of stem cells from international registry. During the Covid19 pandemic, the SCT activity of centers decreased according to the national, and international guidelines. The study revealed an organized, and multidirectional capacity of the registry and also the adaptation to unpredicted conditions such as pandemic. On the other hand, there is a need for more effective strategies for donor recruitment and retention programme.

EFI region 8 Balkan external proficiency testing for HLA typing: Results of 15 years.

Istanbul Medical Faculty, Department of Medical Biology, as the first EFI-accredited HLA laboratory in Turkey since 1999 has been organizing both national and international quality control tests for HLA typing under the banner of the "Balkan external proficiency testing (BEPT)" encompassing countries from EFI regions 8. The first round of BEPT in 2004 was organized for low-resolution HLA-A,-B typing by molecular methods and 12 centres participated in the exercise. In 2005, low-resolution HLA-DR typing was added, and in 2007, low-resolution HLA-C and DQ typing were added to the exercise and 28 centres participated. In 2015, high-resolution (four digits or higher typing) HLA-A,-B,-C,-DR and -DQ typing added to the exercise and 40 centres participated. In the last 2 years, 2017 and 2018, the number of participating centres increased to 48 and 52, respectively. When the distribution of low-resolution typing methods applied in the exercises were investigated, it was found that 82% of the centres in the first round of BEPT in 2004 used PCR-SSP, whereas in the last round in 2018, 26% of the centres preferred SSP, while the rest used SSO (50%) or SSP and SSO (24%) methods. Methods for high-resolution typing were SBT (41%), NGS (18%), SSO (18%), SSP (6%), SBT + NGS (6%) and SBT + SSP (11%). In 15th trial for BEPT, nomenclature mistake rate was 12%, and erroneous result rate was 6% for HLA samples. The most common mistakes in the exercises were nomenclature mistakes. The EFI Standards Version 7.0 is stated "HLA type can be reported using a hyphen if homozygosity is not proven by family studies." In order to provide standard results among HLA laboratories and since accurate HLA typing leads to significant increase of graft and patient survival, quality control exercises should be performed in all HLA-based tests periodically.

Cytokine Polymorphism and HLA Genotyping in Patients with Temporal Lobe Epilepsy Related to Hippocampal Sclerosis.

OBJECTIVE: Hippocampal sclerosis (HS) is the most common pathological substrate associated with mesial temporal lobe epilepsy (MTLE), where inflammatory processes are known to play an increasingly important role in the pathogenesis. To further investigate the role of the immune system, both cytokine gene polymorphisms and human leukocyte antigen (HLA) genotyping in patients with MTLE-HS were investigated. METHODS: The DNA samples of 100 patients with MTLE-HS and 201 healthy individuals were genotyped for cytokines (IL-6,IL-10, TNF-α, TGF-ß1 and IFN-γ) and HLA using polymerase chain reaction (PCR)-SSP and SSO methods. The results were statistically analyzed in patient and healthy control groups and then according to the presence of febrile seizures (FS) in the patient group. RESULTS: Analysis of cytokine genotyping did not reveal any significant difference between patients with MTLE-HS and controls and patients with or without FS. However, the HLA DRB1*13 allele was found to be more frequent in the patient population after Bonferroni correction. CONCLUSION: This study suggests the possible role of HLA in the pathogenesis of MTLE-HS, although it failed to show any relationship with the cytokine system. However, data regarding the role of HLA are still lacking, and further studies are necessary to verify our results.