Mononuclear cells （MNCs） isolated from peripheral blood by density gradient centrifugation were plated on human fibronectin-coated culture plates and cultured in EGM-2 medium. Attached spindle-shaped cells, reported...Mononuclear cells （MNCs） isolated from peripheral blood by density gradient centrifugation were plated on human fibronectin-coated culture plates and cultured in EGM-2 medium. Attached spindle-shaped cells, reported as endothelial progenitor cells （EPCs） by some investigators, had elongated from adherent round cells, but had not proliferated from a small number of cells as supposed previously. The growth curve of the primary EPCs showed that the cells had little proliferative capacity. Flow cytometry analysis showed that the cells could express some of the endothelial lineage markers, while they could also express CD 14, which is considered a marker of monocyte/macrophage lineages throughout culture. In endothelial function assays, the cells demonstrated a lower level of expression of eNOS than mature endothelial cells in the reverse transcription-polymerase chain reaction and did not show an ability to develop tube-like structures in angiogenesis assay in vitro. In this study, we identified the monocytoid function of EPCs by the combined Dillabeled acetylated low-density lipoprotein （Dil-Ac-LDL） and Indian ink uptake tests. All the cells were double positive for Dil- Ac-LDL and Indian ink uptake at days 4, 14 and 28 of culture, which means the EPCs maintained monocytoid function throughout the culture. Therefore, although adult EPCs from peripheral MNCs have some endothelial lineage properties, they maintain typical monocytic function and have little proliferative capacity.展开更多
The chitinase-like proteins YKL-39 (chitinase 3-like-2) and Chitortriosidase (CHIT-1) are members of the chitinases family. YKL-39 expression has been associated with osteoarthritis, whereas CHIT-1 activity is regarde...The chitinase-like proteins YKL-39 (chitinase 3-like-2) and Chitortriosidase (CHIT-1) are members of the chitinases family. YKL-39 expression has been associated with osteoarthritis, whereas CHIT-1 activity is regarded as a biochemical marker of macrophage activation. So far, the physiological or pathological role of YKL-39 in the inflammation is still poorly understood. We compared YKL-39 and CHIT-1 modulation during monocyte to macrophage transition and polarization. Gene expression analysis was investigated by real-time PCR from mRNA of human monocytes obtained from buffy coat of healthy volunteers, from mRNA of polarized macrophages to classically activated macrophages (or M1), obtained by interferon-γ and lipopolysaccharide exposure, and from mRNA of alternatively activated macrophages (or M2) obtained by interleukin-4 exposure. We demonstrated different variations of YKL-39 and CHIT-1 production during macrophages polarization. CHIT-1 levels gradually increase in the course of the time with a peak of expression between the fifth and the seventh day of culture. In contrast, YKL-39 expression was unaltered in the diverse stage of HMMs differentiation, but increased significantly in M1 polarized macrophages and reverted to base levels in M2 polarized macrophages. These findings indicated that the function of YKL-39 is much more restricted and selective than that exerted by CHIT-1.展开更多
基金the National Natural Science Foundation of China （Nos. 30170932 , 30371411） the Foundation for Excellent Young Scholar （No. 30125039）.
文摘Mononuclear cells （MNCs） isolated from peripheral blood by density gradient centrifugation were plated on human fibronectin-coated culture plates and cultured in EGM-2 medium. Attached spindle-shaped cells, reported as endothelial progenitor cells （EPCs） by some investigators, had elongated from adherent round cells, but had not proliferated from a small number of cells as supposed previously. The growth curve of the primary EPCs showed that the cells had little proliferative capacity. Flow cytometry analysis showed that the cells could express some of the endothelial lineage markers, while they could also express CD 14, which is considered a marker of monocyte/macrophage lineages throughout culture. In endothelial function assays, the cells demonstrated a lower level of expression of eNOS than mature endothelial cells in the reverse transcription-polymerase chain reaction and did not show an ability to develop tube-like structures in angiogenesis assay in vitro. In this study, we identified the monocytoid function of EPCs by the combined Dillabeled acetylated low-density lipoprotein （Dil-Ac-LDL） and Indian ink uptake tests. All the cells were double positive for Dil- Ac-LDL and Indian ink uptake at days 4, 14 and 28 of culture, which means the EPCs maintained monocytoid function throughout the culture. Therefore, although adult EPCs from peripheral MNCs have some endothelial lineage properties, they maintain typical monocytic function and have little proliferative capacity.
文摘The chitinase-like proteins YKL-39 (chitinase 3-like-2) and Chitortriosidase (CHIT-1) are members of the chitinases family. YKL-39 expression has been associated with osteoarthritis, whereas CHIT-1 activity is regarded as a biochemical marker of macrophage activation. So far, the physiological or pathological role of YKL-39 in the inflammation is still poorly understood. We compared YKL-39 and CHIT-1 modulation during monocyte to macrophage transition and polarization. Gene expression analysis was investigated by real-time PCR from mRNA of human monocytes obtained from buffy coat of healthy volunteers, from mRNA of polarized macrophages to classically activated macrophages (or M1), obtained by interferon-γ and lipopolysaccharide exposure, and from mRNA of alternatively activated macrophages (or M2) obtained by interleukin-4 exposure. We demonstrated different variations of YKL-39 and CHIT-1 production during macrophages polarization. CHIT-1 levels gradually increase in the course of the time with a peak of expression between the fifth and the seventh day of culture. In contrast, YKL-39 expression was unaltered in the diverse stage of HMMs differentiation, but increased significantly in M1 polarized macrophages and reverted to base levels in M2 polarized macrophages. These findings indicated that the function of YKL-39 is much more restricted and selective than that exerted by CHIT-1.
