Characterizations of cd34+ stem cells from hematopoietic stem cells (hscs)

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Author: 
Ahmad Fazilat
Abstract: 

Stem cells are unspecialized cells in the human body that are capable of becoming specialized cells, each with new specialized cell functions. Hematopoietic stem cells (HSCs) originated from mesodermal hemangioblast cells which have an ability to self-renew and multipotent nature. These HSCs usually express CD34+ and CD133+ glycoproteins on their surface and antibodies raised against these surface glycoproteins are the best exploited matter in the purification steps. CD34+ is a cluster of differentiation molecule presents on certain cells within the human body. The present research mainly focused on characterization of CD34+ stem cells from HSCs, which is in associated with diagnose of varius tumors such as alveolar soft part sarcoma, AML-M7 and preB-ALL. CD34 cells were maintained in the laboratory under appropriate conditions. The cells were cultured in DMEM with 95% humidity and 5% CO2. They showed round shaped actively dividing state under daily observations. These cells were stained with Leishman-Giemsa (LG) stain showed nucleated structures with no contamination with other morphologically dissimilar structures. The expression of CD34 surface glycoprotein in the growing cells was confirmed by immunocytochemistry (ICC). For detection of CD34 marker on the cultured cells, the Anti CD34 antibody was used and probed against anti-mouse HRPO sterpavidin conjugate as secondary antibody. The secondary antibody when added with developer, DAB-H2O2 given brown Colour precipitate colony forming unit assay showed efficiency that CD34+ stem cells can be determined based on number of colonies which emerge. This is based on the ability of cells to grow unattached and to remain suspended in agar. The results showed the presence of CD34+marker where the AntiCD34 antibody binds specifically to the CD34+ marker present on the cell surface of hematopoietic stem cells.

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