Abstract
BackgroundThe main morphological features of primitive cells, such as stem and progenitor cells, are that these cells consists only one nucleus. The main purpose of this study was to determine the differentiation capacities of stem and progenitor cells. This study was performed using mononucleated cells originated from murine peripheral blood and MC3T3-E1 cells. Three approaches were used to determine their differentiation capacities: 1) Biochemical assays, 2) Gene expression analysis, and 3) Morphological observations.ResultsWe found that both cells were able to differentiate into mature osteoblasts, as assayed by ALP activity. RT-PCR analysis showed the activation of the Opn gene after osteoblast differentiation. Morphological observations of both cells revealed the formation of black or dark-brown nodules after von Kossa staining. Nevertheless, only mononucleated cells showed the significant increase in TRAP activity characteristic of mature osteoclasts. The osteoclast-specific CatK gene was only upregulated in mononucleated cells. Morphological observations indicated the existence of multinucleated osteoclasts. Sca-1 was activated only in undifferentiated mononucleated cells, indicating that the cells were hematopoietic stem cells. In both cell lines, the housekeeping Gapdh gene was activated before and after differentiation.ConclusionThe isolated mononucleated cells were able to differentiate into both osteoblasts and osteoclasts; indicating that they are stem cells. On the other hand, MC3T3-E1 cells can only differentiate into osteoblasts; a characteristic of progenitor cells.
Highlights
The main morphological features of primitive cells, such as stem and progenitor cells, are that these cells consists only one nucleus
alkaline phosphatase (ALP) is a biochemical marker of osteoblasts, while tartrate-resistant acid phosphatase (TRAP) is a biochemical marker of osteoclasts, as the ALP and TRAP enzymes are secreted during osteoblast and osteoclast differentiation, respectively
ALP activity in mononucleated and MC3T3-E1 cells gradually increased from day 3 to day 14 of osteoblast differentiation
Summary
The main morphological features of primitive cells, such as stem and progenitor cells, are that these cells consists only one nucleus. Stem cell research has spread into many fields of study, indicating that it represents an area with great scientific and therapeutic promise Their unique ability to self-renew indefinitely and to differentiate into multiple cell types can make them useful to elucidate normal cellular processes as well as to understand their mechanisms [1]. For example Seaberg and van der Kooy [3] had stated that some researchers exclude the characteristic of stem cells plasticity as one of the novel property of stem cells They suggested that there are differences in the biological marker of these two cell types, i.e., hematopoietic stem cells and progenitor cells detected in both in vivo and in vitro studies. A few potential biological markers, such as Thy-1 and Sca-1 have been used to prove the difference for respective cells
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