Human CD34(+) hematopoietic cells, a distinctive cell population containing hematopoietic stem/progenitor cells (HSPC), have the capability to highly self-renewal, differentiation into all lineages of committed progenitor cells and reconstitution of both long-term hematopoiesis and immunefunctions after transplantation. CD34(+) hematopoietic cells from bone marrow (BM) recently have been employed for treating neoplastic and genetic disorders. This study was aimed to investigate membrane surface ultrastructures of bone marrow CD34(+) cell from mormal persons and leukemia patients and to compare their morphologic differences by using atomic force microscope (AFM). BM was collected from 5 normal donors and 6 leukaemia patients. All samples were layered on Ficoll-Paque gradients (specific gravity 1.077 g/ml) to separate the mononuclear cells. After that CD34(+) cells were purified by immuno-magnetic bead separation and evaluated with a FACS Calibur, these cells were detected by AFM of tapping mode inair. At lest 20 cells per samples were observed. The results showed that most of CD34(+) hematopoietic cells were like circle plate, the diameter was 10 - 14 microm. The surface of CD34(+) hematopoietic cell membrane was comparatively complex. The surface of CD34(+) hematopoietic cell membrane appeared as granular, with packed particles. With the region analysis function of IP2.1 software, the region of 2 microm x 2 microm was selected and four parameters of the surface (maximum peak-to-valley distance, average roughness, root-mean-squared roughness and mean height) were measured. Values of the 4 parameters showed that the characteristic parameters of CD34(+) HSPC from leukaemia were higher than that from normal person. It is concluded that AFM has specific advantages in analyzing cell membrane in the nanometer level and can gain more information. With the help of analysis software, AFM can be a helpful tool for fast leukaemic diagnosis and CD34(+) hematopoietic cells selection.
Antigens, CD34 ; analysis ; Bone Marrow Cells ; cytology ; immunology ; ultrastructure ; Cell Membrane ; ultrastructure ; Hematopoietic Stem Cells ; cytology ; immunology ; ultrastructure ; Humans ; Microscopy, Atomic Force

In order to investigate the ultrastructural features of malignant T cell (MTC) in bona marrow aspirate (BMA) from patients with T Cell Lymphoma, the antigen expression of MTC was analyzed by flow cytometry, and the ultrastructural features of MTC in BMA from 13 T-cell lymphoma patients with bone marrow involvement (BMI) were observed by transmission electron microscopy. The results indicated that the sizes of MTC were uneven in every patient and their diameter were between 12 and 28 microm, in 6 out of 13 cases sizes of MTC were slightly uneven but in 7/13 cases sizes of MTC were significantly uneven. The heterochromatin of MTC was less than that of normal T cell and nucleolus diameter was from 2 to 8 microm in all cases. The nuclear contour of MTC was strikingly irregular in 10 out of 13 cases. The MTC had plenty of cytoplasm in 8 out of 13 cases and displayed many microvilli or processes on MTC surface in 7 out of 13 cases, while MTC in 6 out of 13 cases contained more Golgi's apparatuses, secretary vacuoles, dense granules and intermediate filaments. In 8 out of 13 cases mitochondria apparently swelled. It is concluded that the size of MTC increase unevenly in all patients. MTC nuclear contour in most cases is irregular by folding, indenting, and twisting, which often correlated with arising of paranuclear intermediate filaments. Processes and microvilli on surface and Golgi's apparatus, secretary vesicles, dense granules as well as intermediate filament in cytoplasm of MTC develop synchronously, meanwhile, mitochondria of MTC strikingly swell in most cases.
Adolescent ; Adult ; Aged ; Bone Marrow Cells ; ultrastructure ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Lymphoma, T-Cell ; pathology ; ultrastructure ; Male ; Middle Aged ; Neoplasm Invasiveness ; T-Lymphocytes ; ultrastructure

OBJECTIVE: To observe the repairing effects of bone marrow transplantation with nerve tissue committed stem cell (NTCSCs) on experimental rats with injury of noise-induced hearing loss. METHOD: Guinea pigs were randomly divided into control group, noise exposure group and the transplanting group. A week after white noise exposure of 110 dB, NTCSCs and PBS were injected into guinea pigs of the noise exposure group and the transplanting group respectively. One week after noise exposure to four weeks continuous administration. ABR thresholds were measured respectively prior to the experiment, 1 week post-noise,1, 2 and 4 weeks post-drugs, The changes of cochlea hair cells were also observed by a scan electron microscope (SEM). RESULT: The ABR threshold shifts in the transplanting group were significantly fewer than that in the noise exposure group. SEM showed that hear hair of the inner and outer hair cells in noise exposure group displayed mess, fusion and imperfections. In the transplanting treatment group, the hair cells displayed slight pathological changes, there wasn't significant differents comparied with normal group. The number of OHCs were relatively stable in the normal group, while the obvious OHC loss was observed in other groups. There was significant difference among the three groups, however, the OHC loss in the transplanting group was no significantly different to that in the noise exposure (P > 0.05). CONCLUSION The bone marrow NTCSCs which had been transplanted to rat cochlea could reduce the damage of the noise on the hair cell, and thus played a role in repairing the damage of auditory nerve.
Animals ; Bone Marrow Cells ; Bone Marrow Transplantation ; Cochlea ; Guinea Pigs ; Hair Cells, Auditory, Outer ; pathology ; ultrastructure ; Hearing Loss, Noise-Induced ; therapy ; Noise ; adverse effects ; Rats ; Stem Cell Transplantation

The effect of four alkylating agents (MMS, EMs, DMN, DEM), under various con centrations on mouse bone marrow erythrocytes, were studied by means of the micronucleus test. The results obtained were as follows: 1) The lethal doses on mice were MMS = 130 mg/kg/bw, EMS = 300 mg/kg/bw, DMN = 50 mg/kg/bw and DEN = 70 mg/kg/bw. 2) Micronuclei were easily seen and in different controls the micronulei were found a little over 0.1%. 3) The dose-effect relationship was obtained. In the MMS and EMS treated groups, incidences of micronulei were 0.45 to 2.56% and 0.4 to 2.1% respectively. 4) In the DMN and DEN treated groups, incidences varied between 0.15 to 0.90 % and 0.2 to 1.02% respectively. 5) Four alkylating agents were compared and discussed with respect to micro nucleus production from chromosomal aberrations.
Animal ; Bone Marrow/ultrastructure ; Carcinogens/pharmacology* ; Cell Nucleus/drug effects* ; Chromosome Aberrations* ; Erythrocytes/ultrastructure ; Female ; Mesylates/pharmacology* ; Mice ; Mutagens/pharmacology ; Nitrosamines/pharmacology*

OBJECTIVETo further investigate the osteogenic potential of rabbit marrow stromal stem cells cultured in vitro.

METHODSRabbit marrow stromal stem cells were isolated by density gradient centrifugation method and amplified in the flasks, using the osteogenic inducing conditions (OGC) as the culture media. The osteogenic potential of marrow stromal stem cells were investigated by means of bone-seeking fluorescence (tetracycline) labelling, Alizarin red S (ARS) staining, Alcian blue-Sirius red (AS) staining, and scanning electron microscope.

RESULTSAfter being passaged, the marrow stromal stem cells increased in number, became confluent and formed multi-layer structure. The stromal stem cells excreted innumerable tiny granules, heaping up on the cell body and merging gradually into foggy substances. These foggy substances kept on enlarging and formed round, oval, or flake-like nodules. These nodules revealed bright golden yellow fluorescence under fluorescence microscope when labelled with tetracycline. Histochemical study with specific new bone staining with ARS revealed positive calcium reaction, both denoting that they were newly formed bone tissues. After they were stained with AS, collagen and acid mucopolysaccharide were shown. Under scanning electron microscope, three types of cells with different configurations were found. They were globular cells, spindle-shaped cells and polygonal or polygonal cells. Granules were excreted from the cells and heaped up on the cell body. Needle-shaped and irregularly rectangular crystals also appeared and agglomerated with the granules to form nodules and trabecula-like or flake-like structures.

CONCLUSIONSSequence of events of bone formation by rabbit marrow stromal stem cells cultured in vitro is fully depicted and confirmed, which provides the foundation for further investigating the mechanisms of osteoblast differentiation from marrow stromal stem cells and the possible application in orthopaedics.

Animals ; Bone Marrow Cells ; pathology ; ultrastructure ; Cells, Cultured ; Immunohistochemistry ; In Vitro Techniques ; Male ; Microscopy, Electron, Scanning ; Models, Animal ; Osteogenesis ; physiology ; Rabbits ; Sensitivity and Specificity ; Stromal Cells ; physiology ; ultrastructure

The study was aimed to investigate the ultranstructural feature and diagnostic criteria of congenital dyserythropoietic anemia-type I (CDA-type I). Nucleated red cells in bone marrow from two patients with CDA-type I were analyzed by transmission electron microscopy (TEM). The results indicated that the erythropoietic/granulopoietic ratio was markedly increased with megaloblastic morphology in all stage of erythrocyte. Most proerythroblast showed of irregular nuclei, while the Swiss-cheese-appearance of the heterochromatin was usually found in basophilic and polychromatic erythroblast. About half of orthochromatic erythroblast illustrated karyolysis and karyorrhexis. Some orthochromatic erythroblast exhibited karyolysis and plasmolysis simultaneously. The inter-nuclear chromatin bridge between separated erythroblasts was seldom found by TEM. The nuclear membrane and rough endoplasmic reticulum were destructed at all stage of erythrocytes in different degree. In conclusion, the megaloblastic erythrosis was the main characteristic of CDA-type I, and then nuclear membrane disruption in polychromatic erythroblast and karyolysis or karyorrhexis in orthochromatic erythroblast. The universal breakdown of cytoplasm membranous system was fundamental pathogenesis of CDA-type I.
Anemia, Dyserythropoietic, Congenital ; blood ; pathology ; Bone Marrow Examination ; Erythroblasts ; ultrastructure ; Erythrocytes ; ultrastructure ; Female ; Humans ; Infant ; Iron ; blood ; Male ; Microscopy, Electron, Transmission

Light and electron microscopic studies of megakaryocytes in bone marrow from 14 cases of Korean epidemic hemorrhagic fever were made, together with clinical manifestations, laboratory findings, and changes in other marrow elements. The number of megakaryocytes increased in various degree from patient to patient, with a shift to the left of maturation gradation. Cytoplasms were markedly smaller but granularity was fairly good. The most constant finding was marked decrease or complete absence of platelet formation at the periphery of the cytoplasm. Ultrastructurally, there was a marked decrease of the cisternal system in the cytoplasm and lack of cleavage line. No virus particles or active degenerative process was noted. These changes of the megakaryocytes are similar to those reported in idiopathic thrombocytopenic purpura, indicating that megakaryocytic changes in Korean epidemic hemorrhagic fever are likely secondary to thrombocytopenia which was caused by other Mechanism.
Adult ; Aged ; Bone Marrow/pathology ; Female ; Hemorrhagic Fever with Renal Syndrome/pathology* ; Human ; Male ; Megakaryocytes/ultrastructure* ; Middle Age

The aim of study was to investigate the relationship of anemia and neutropenia with ultrastructural abnormalities of erythroblasts and young neutrophils in bone marrow of patients with myelodysplastic syndrome (MDS). Anemia parameters and peripheral neutrophil amount of 74 patients with MDS were measured by automatic hemocyte analyzer. According to Hb value and neutropenia degree, MDS patients were divided into 4 groups: normal, mild, middle and severe anemia or neutropenia. The morbid rate and apoptosis rate of erythroblasts and young neutrophils in bone marrow were measured by transmission electron microscopy (TEM). The results indicated that 68 out of 74 patients were consistent with anemia diagnostic criteria, and 51 out of 68 patients were with neutrocytopenia. TEM showed different abnormal features of erythroblasts and young neutrophils in all patients. The morbid rates of erythroblasts in normal, mild, middle and severe anemia groups were 37 ± 14.7%, 24 ± 9%, 32 ± 16% and 34 ± 21% respectively, while apoptotic rates of erythroblasts in normal, mild, middle and severe anemia groups were 2.25 ± 1.03%, 4.43 ± 2.60%, 8.78 ± 4.04% and 11.67 ± 4.57% respectively. The morbid rate and apoptotic rate of erythroblasts were correlated negatively with Hb and HCT value (p < 0.05). The apoptotic rates of bone marrow young neutrophils in 4 groups with different degree of neutropenia were 6.00 ± 2.67%, 9.50 ± 4.42%, 13.00 ± 3.54% and 17.00 ± 2.39%, which correlated negatively with peripheral neutrophil quantity (p < 0.01). Morbid rates of neutrophils in normal, mild, middle and severe anemia groups were 12.25 ± 16.31%, 13.5 ± 10.01%, 23 ± 8.59% and 51.67 ± 19.67% respectively, which positively correlated with its apoptotic rates (p < 0.01). It is concluded that anemia and neutropenia in patient with MDS are correlated with apoptosis and morbid rate of erythroblasts and young neutrophils in bone marrow, which may result in ineffective hematopoiesis.
Adult ; Anemia ; complications ; pathology ; Apoptosis ; Bone Marrow Cells ; cytology ; ultrastructure ; Female ; Humans ; Male ; Myelodysplastic Syndromes ; complications ; pathology ; Neutropenia ; complications ; pathology

OBJECTIVETo observe the ultrastructure of bone marrow stromal cells (BMSCs) cultured in coralline hydroxyapatite (CHA) and evaluate their biocompatibility.

METHODSBMSCs isolated from dogs were cultured with CHA as the scaffold, and the morphologies of the cells were observed with phase-contrast microscope and scanning electron microscope.

RESULTS AND CONCLUSIONBMSCs grew well with good attachment to the CHA scaffold and performed normal function, demonstrating CHA as one of useful biocarrier materials for bone tissue engineering.

Animals ; Bone Marrow Cells ; cytology ; ultrastructure ; Bone Substitutes ; chemistry ; Cell Culture Techniques ; Cells, Cultured ; Ceramics ; chemistry ; Dogs ; Hydroxyapatites ; chemistry ; Male ; Microscopy, Electron, Scanning ; Microscopy, Phase-Contrast ; Stromal Cells ; cytology ; ultrastructure ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

A rare case of Behcet's disease associated with myelodysplastic syndrome (MDS) is described. A 50-year-old Korean female suffering recurrent oral ulcer, genital ulcer, fatigue, arthralgia in both knees and fever was diagnosed as Behcet's disease. The findings of bone marrow aspirates were consistent with refractory anemia, a subtype of myelodysplastic syndrome. Chromosomal analysis of bone marrow cells revealed 46,XX,-8,-20,+der(8)t(8;20)(p23;p10),+der(8) t(8;20)(p23;q10)[30]. The chromosomal changes found in this patient were different from those of previous reports, which mostly revealed trisomy 8. If anemia, low reticulocyte count and dyspoietic cells are sustained in Behcet's disease, physicians should be alert to the possibility of MDS with aberration in chromosome 8 and perform a bone marrow study for the proper diagnosis and treatment of the disease. We presented a case of Behcet's disease associated with MDS, which is the first Korean case.
Anemia/genetics ; Behcet's Syndrome/genetics* ; Behcet's Syndrome/diagnosis ; Bone Marrow Cells/ultrastructure ; Bone Marrow Cells/pathology ; Case Report ; Chromosome Aberrations ; Female ; Histocytochemistry ; Human ; Korea ; Middle Age ; Myelodysplastic Syndromes/genetics*