Abstract. The aim of this study was to investigate changes in primitive hematopoietic cells through CD38 expression, identify the stage at which erythrocyte differentiation CD38 gains activity and the effects of serum factors on this expression by establishing a hematopoietic stem cell system in the erythroid development process. Using an immunomagnetic labeling and separation technique, CD34 + cells were selected from cord blood. The CD34 + cells were cultured in a 2 mM L-glutamine-enriched medium containing erythropoietin (Epo), penicillin-streptomycin and stem cell factor (SCF), and were incubated in 5% CO 2 at 37˚C. In erythroid development pathways following CD38 expression, primitive/progenitor human hematopoietic cells obtained from cord blood were assessed through the erythroid development process in a serum-free medium in the presence of proper SCF and Epo. At the end of the 26-day process, using staining with a Megacult-c staining kit, it was determined that progenitor cells nucleate and differentiate into erythroid cell lines of 8-10 µm. During the course of this process, we analyzed increases over time in NAD glycohydrolase activity rates using the supernatant liquid samples. Results of co-culture experiments in cell culture studies showed that the stimulating effects of CD38 expression originate from specific serum factors. CD38 expression has been shown to occur at hematopoietic cell sources as well as at a number of differentiation levels. In the proliferation process the possible induction of CD38 through specific serum factors leads us to conclude that it may be involved in proliferation with a physiological task or that it may be involved in an event, such as an apoptotic process.
Abstract. Erythrocyte and lymphocyte NAD + glycohydrolase levels were previously found to be elevated in cancer patients. These results were confirmed in an animal model. The administration of live Ehrlich ascites tumor cells to BALB/c mice led to increases in erythrocyte and lymphocyte NAD + glycohydrolase, along with tumor development. Serum samples, ascites fluid from mice with developed tumors, serum samples from cancer patients and Ehrlich cell supernatants had a similar stimulatory effect when administered to mice or when incubated with peripheric lymphocytes in culture. These increases were accompanied by the appearance of an anti-CD38 reactive band of 45 kDa in SDS-PAGE/Western blot analyses of erythrocyte ghost and lymphocyte membrane proteins. The results, supported by flow cytometry data, support previous clinical findings that an enhancement in CD38 expression occurs in the hematopoietic system during proliferative processes. Moreover, they suggest that CD38 expression is triggered at least in part by a certain cytokine(s) secreted by cancer cells. Finally, the results emphasize the prospective use of CD38 expression as a marker of tumor development and progression.
Objective:The preferred method for the purification of hematopoietic stem cells (HSCs), which can easily differentiate into blood cells, is the selection of CD34 + cells by flow cytometry or magnetic discrimination. Methods are needed that effectively determine human stem cells without entirely relying on phenotypic cell surface markers. One of these methods is cytosolic aldehyde dehydrogenase (ALDH), which plays a role in retinoid metabolism and resistance of HSCs to alkylating agents such as cyclophosphamide. The prospective isolation of human hematopoietic stem and progenitor cells with different functions, as well as surface markers, are also recommended by ALDH and flow cytometry. In the above findings, ALDH + cells can be isolated from other cells and the most primitive hematopoietic stem cells can be isolated and used in clinical applications. Materials and methods: In our study, we separated the CD34 surface marker cells from cord blood using magnetic nanoparticles and then purified the cells with ALDH activity and non-CD38 surface antigen by flow cytometer. We visualized these cell groups with fluorescence microscopy. Results: We observed that the majority of CD34 + cells were ALDH + in fluorescence microscopy and flow cytometry (FACS) analysis of CD34 + cells purified from cord blood as a result of ALDH-FITC markings.
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