Bone Marrow Cells ; cytology ; Fibroblasts ; cytology ; metabolism ; Fibrosis ; Humans ; Myocardium ; pathology

The genesis and development of leukemia not only associate to intrinsic factors, but also relate with the fibrous hyperplasia in the bone marrow. This review mainly focuses on the interaction between fiber-producing cells and leukemia cells, the relationship between fibrous hyperplasia and prognosis of leukemia, the regulation of TGF-beta, PDGF and other cytokines, the underlying mechanism of fibrous hyperplasia so as to explore the potential therapeutic targets for improving the prognosis of leukemia.
Bone Marrow ; pathology ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cytokines ; metabolism ; Fibroblasts ; cytology ; Humans ; Leukemia ; pathology ; Platelet-Derived Growth Factor ; metabolism ; Transforming Growth Factor beta ; metabolism

This study was purposed to investigate the connexin 43 (Cx43) expression level in acute leukemia bone marrow stromal cells (ABMSCs) and normal bone marrow stromal cells (NBMSCs), and to explore the difference in communicating functions between these cells. The Cx43 expression levels of ABMSCs and NBMSCs were detected by using immunohistochemistry and computer gray scale assay, and the difference of gap junction intercellular communication (GJIC) was examined through dry transfer technique. The results showed that expression level of Cx43 in ABMSCs was lower than that in NBMSCs and its function of GJIC in ABMSCs was also weaker than that in NBMSCs. It is concluded that cell-cell communication function is lowered in ABMSCs.
Acute Disease ; Bone Marrow Cells ; cytology ; metabolism ; pathology ; Cell Communication ; physiology ; Connexin 43 ; metabolism ; Gap Junctions ; metabolism ; Humans ; Leukemia ; metabolism ; Stromal Cells ; cytology ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo explore the autophagy activity of CD34+ cells in bone marrow of MDS patients and its clinical significance.

METHODSThe activity of autophagy in bone marrow CD34+ cells from 20 MDS patients, 20 non-malignant anemia patients and 5 AML patients admitted in our hospital from October 2012 to March 2014 was detected by flow cytometry (FCM).

RESULTSThe autophagy activity in low risk MDS patients and non-malignant anemia patients were both significantly higher than that in both high risk MDS and AML patients (P<0.05), and more interestingly, the autophagy activity in MDS negatively correlated with World Health Organization classification-based prognostic system (WPSS) score (r=-0.877) .

CONCLUSIONThe autophagy activity CD34+ cells in the patients with MDS is higher than that in AML patients, and negatively correlated with WPSS scores, indicating that the decrease of autophagy activity maybe accelerate the genesis and development of MDS and relate with the prognosis of MDS patients.

Antigens, CD34 ; metabolism ; Autophagy ; Bone Marrow Cells ; cytology ; pathology ; Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; pathology ; Myelodysplastic Syndromes ; pathology ; Prognosis

This study was aimed to investigate the conditions of culturing in vitro mesenchymal stem cells (MSCs) derived from bone marrow of children with acute leukemia and the biological characteristics of MSCs from leukemia children. The bone marrow MSCs of acute leukemia children were isolated by density gradient centrifugation combined with adherent segregating method and cultured in DMEM/F12. The morphology of Wright stained MSCs was observed under inverted microscope. Cell surface markers were analyzed with flow cytometry. The growth characteristic features of cultured MSCs was measured with MTT method. Induced adipogenic and osteogenic differentiation of MSCs in appropriate induction media was observed. The results indicated that BM-MSCs of acute leukemia children could be successfully cultured in vitro in appropriate conditions. At 24 hours of culture the MSCs began to adhere to wall, grew in colony and appeared in different shapes. As the culture lasted, the MSCs proliferated continuously and shaped in fusiform. After 2 - 3 weeks of culture, MSCs covered the bottom of culture flask. The analysis of growth feature showed that MSCs were in latency for 3 days, and then entered into growth period. After 8 days of culture the growth of MSCs showed to be in plateau stage. The shape of MSCs in 1st and 2nd generation showed to be heterogeneous but the 3rd generation to be homogeneous with long-fusiform. Cells were arranged in shape of whirlpool or radiation. The surface marker analysis showed that the MSCs were positive for CD105, CD29, CD13, but negative for CD34, CD45, CD14 and HLA-DR. The MSCs from leukemia children could be induced into adipocytes and osteocytes in appropriate conditions. It is concluded that (1) MSCs derived from children with acute leukemia can be successfully cultured and passaged in vitro; (2) MSCs from leukemia children not received chemotherapy are more successfully cultured in vitro than those received chemotherapy; (3) the common biological characteristics of MSCs from children with acute leukemia are same as the MSCs from healthy person.
Adolescent ; Bone Marrow ; pathology ; Bone Marrow Cells ; cytology ; metabolism ; Cell Culture Techniques ; Child ; Child, Preschool ; Female ; Humans ; Leukemia ; metabolism ; pathology ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Tumor Cells, Cultured ; Young Adult

We hypothesized that the formation and differentialtion of osteoclasts are accelerated and the potential of bone resorption is increased in the hemiplegic bone marrow in the early stage of stroke. We randomly divided white female Sprague-Dawley (SD) rats (n = 30) into two groups, stroke (n = 15) and sham group (n = 15). On the 7th day after stroke, after cutting away the epiphyses of the femurs and tibias, diaphyseal channels were flushed using alpha-minimum essential medium (alpha-MEM) and bone marrow cells were collected. Bone marrow stem cells, which were extracted from the femur and tibia, were cultured on the 7th day after middle cerebral artery occlusion. We then estimated the ratio of non-adherent cells to total bone marrow cells that included osteoclast precursor cells. After culturing these cells separately, cells that tested positive on the tartrate resistant acid phosphatase (TRAP) were counted and bone resorption was evaluated by using the OAAS(TM) plate. In comparison to the control group, the stroke group showed a higher increase of non-adherent cells in the hemiplegic side bone marrow. In addition, after the primary culture, the stroke group showed an increased number of TRAP positive cells and a higher degree of bone resorption estimated by OAAS(TM) plate. As a result, osteoclastogenesis and osteoclast differentiation are accelerated and the potential of bone resorption is increased in the hemiplegic bone marrow and these changes are detected as early as within the first week after middle cerebral artery occlusion in SD rats.
Animals ; Bone Marrow Cells/cytology/drug effects ; Bone Resorption/*physiopathology ; Cell Differentiation ; Cell Separation ; Cells, Cultured ; Female ; Femur/cytology ; Osteoclasts/cytology ; Rats ; Rats, Sprague-Dawley ; Stem Cells/cytology/metabolism ; Stroke/*metabolism/pathology ; Tartrates/pharmacology ; Tibia/cytology

OBJECTIVETo induce primary chronic myeloid leukemia (CML) cells into dendritic cells (DCs).

METHODSBone marrow mononuclear cells (MNCs) were isolated from 13 CML patients and peripheral blood MNCs from 5 healthy donors. The isolated MNCs were co-cultured with rhGM-CSF 1,000 U/ml, rhIL- 4,500 U/ml and TNF-alpha 50 U/ml for 10 days. The morphological features were observed by Wright's staining,inverted microscope and electron microscope. CD(80), CD(86), CD(83), CD(1a) and HLA-DR expression were assayed by flow cytometry, cytogenetic analysis was performed by fluorescence in-situ hybridization(FISH). The concentration of IL-12 was measured by ELISA and the function of antigen presenting was tested by mixed lymphocyte reaction (MLR).

RESULTAfter being cultured with cytokines, the typical dendritic appearance with delicate membrane projections was observed. The CD(80), CD(86), CD(83), CD(1a) and HLA-DR markers and capacity of stimulating allogeneic T cells were upregulated significantly. FISH confirmed that the DCs were generated from leukemic origin and CML DCs could secrete higher level of IL-12 than CML MNCs. There were no differences in morphology and immunophenotype expression between DCs derived from CML and those from normal individuals. However, DCs from CML patients displayed weaker activity than that of normal individuals when tested in MLR.

CONCLUSIONCML cells could be induced into leukemia-DCs by co-culture with cytokines.

Bone Marrow Cells ; immunology ; pathology ; Cell Differentiation ; Dendritic Cells ; cytology ; immunology ; Humans ; Interleukin-12 ; metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; pathology ; Tumor Cells, Cultured

To investigate the effects of hypoxia on the apoptosis and glucose intake of mesenchymal stem cells (MSCs), MSCs derived from bone marrow of rats were incubated in the atmosphere of 1% O(2) for a series of time points and their glucose-intaking capacity, ultrastructural changes and apoptotic proportions were analyzed by (3)H-labeling assay, electron microscopy and flow cytometry, respectively. The results showed that the cultured cells took the fibroblast-like morphology and could be induced into osteoblasts and adipocytes under appropriate conditions. The proportions of apoptotic cells after hypoxia treatment for 1, 4 and 8 hours were 13.7 +/- 2.26%, 14.1 +/- 2.78% and 14.7 +/- 4.01%, respectively, all of which were significantly higher than that observed in normoxic counterparts (0.09 +/- 2.03%, p < 0.05). Also, cell death occurred after hypoxia treatment and the death rates were 3.11 +/- 2.14%, 4.72 +/- 2.05% and 4.91 +/- 3.72% for 1, 4 and 8 hours incubation respectively. Under hypoxia culture in vitro, cell membrane microvillus began to fall off and the mitochondrias became swelling at 1 hour, and the above changes increasingly aggravated along with hypoxia time prolongation. The (3)H-glucose intaking ratios of MSCs at different hypoxia time points significantly decreased than those in normoxic cells (p < 0.01). It is concluded that the acute hypoxia can induce down-regulation of glucose-intaking capacity, ultrastructural changes and apoptosis of MSCs.
Animals ; Apoptosis ; physiology ; Bone Marrow Cells ; cytology ; Cell Hypoxia ; Cells, Cultured ; Female ; Glucose ; metabolism ; Male ; Mesenchymal Stromal Cells ; metabolism ; pathology ; Rats ; Rats, Wistar

OBJECTIVETo construct a co- culture system of mesenchymal stem cells (MSC) and multiple myeloma (MM) cells and investigate the alterations of connexin 43 (CX43) expression and stromal derived growth factor (SDF)- 1α secretion of MSC.

METHODSCX43 expression and SDF- 1α secretion of MM cell lines (RPMI8226) and human primary MM cells were analyzed by western blot and immunofluorescence. Western blot, RT- PCR and immunofluorescence were employed to detect the alterations of CX43 expression and distribution in MSC directly and indirectly co-cultured with myeloma cells. Lucifer yellow dye spread was utilized to evaluate gap junctional intercellular communication (GJIC) between co- cultured MSC. Transwell was applied to study the transmigration of RPMI8266 induced by MSC under the condition of 18α- glycyrrhetinic acid (18α-GA). The level of SDF- 1α was detected by EILSA.

RESULTSRPMI8266, U266 and one-third primary MM cells expressed CX43 at low or moderate levels. CX43 wasn't expressed in XG- 4 and XG- 7 cells but highly expressed in MSC. The expressions of CX43 mRNA of MSC were up- regulated after directly and indirectly co- cultured with RPMI8226, 1.36 and 2.10 times that of MSC cultured alone respectively. Western blot analysis showed that CX43 protein expression of MSC was also up-regulated, mainly distributed in cytoplasm. Lucifer yellow dye spread showed that GJIC was up-regulated in MSC. SDF-1α concentration in supernatant of MSC directly and indirectly co-cultured with RPMI8226 were (373.02±10.11)pg/ml and (309.71±10.71)pg/ml respectively, which were higher than that of MSC cultured alone (237.84±9.23)pg/ml (P<0.01), and could be inhibited by 18α-GA [(237.84±9.23)pg/ml and (94.31±6.44)pg/ml] respectively (P<0.01). 18α-GA could inhibit the transmigration of RPMI8226 induced by MSC, decrease from (8.00±0.67)% to (4.82±0.19)%.

CONCLUSIONCX43 expression of MSC was up-regulated after directly and indirectly co-cultured with MM cells, which could improve the level of SDF-1α secretion of MSC. GJ inhibitor could downregulate SDF-1α secretion of MSC and inhibit the transmigration of MM cells induced by MSC.

Bone Marrow Cells ; cytology ; Cell Line, Tumor ; Chemokine CXCL12 ; secretion ; Coculture Techniques ; Connexin 43 ; secretion ; Humans ; Mesenchymal Stromal Cells ; cytology ; Multiple Myeloma ; metabolism ; pathology

No abstract available.
Acute Disease ; Aged ; Aged, 80 and over ; Bone Marrow Cells/cytology/metabolism/pathology ; Cell Lineage ; Humans ; Immunophenotyping ; Karyotyping ; Leukemia/*genetics/metabolism/pathology ; Male ; *Tetraploidy