Antigens, CD34 ; Cell Count ; methods ; Flow Cytometry ; Humans

OBJECTIVE: To explore the maintaining measures for the vitality of hematopoietic stem cells (HSC) in vitro, so as provide technical support for ultra long distance transport of HSC collected from unrelated donors. METHODS: Peripheral blood hematopoietic stem cells (PBHSC) were treated by different methods according to various groups, then stored at 4 ℃ in the refrigerator. The percentage of CD34 cells, relative cell activity, relative cell proliferation rate, relative colony-forming rate, oxygen fraction and intracellular reactive oxygen species (ROS) were detected at 0, 24, 48 and 72 h after storage of PBHSC respectively. RESULTS: The percentage of CD34 cells during 72 h storage did not altered. Along with the prolonging of storage time, the relative cell activity, relative cell proliferation rate and relative colony-forming rate gradually decreased in untreated PBHSC(control group), the related coefficients were -0.796, -0.883 and -0.815 respectively. Plasma dilution, antioxidants and oxygenation could improve the relative cell activity and relative cell proliferation rate, but oxygenation could decrease the relative colony-forming rate of PBHSC. The combination of 2 or 3 factors showed stronger protection effects on PBHSC. The intracellular level of ROS decreased gradually with the prolonging of storage time. Oxygenation of PBHSC could increase oxygen fraction, and also increase the intracellular level of ROS at the same time. The addition of antioxidants could reduce the level of ROS. CONCLUSION The percentage of CD34 cells can not serve as the indicator of PBHSC vitality. Plasma dilution, oxygenation and antioxidants can increase the survival and viability of PBHSC, but oxygenation can increase the intracellular ROS level and impair colony-forming ability of PBHSC. The combination of multiple factors can maintain the vitality of PBHSC better.
Antigens, CD34 ; Antioxidants ; Hematopoietic Stem Cells ; Reactive Oxygen Species

OBJECTIVES: To observe the changes of the formation time of venous thrombus in rats, and to provide new ideas and methods for the estimation on thrombus formation time of the forensic cases died from thrombosis. METHODS: Totally 80 rats were randomly divided into 10 groups （0 h, 3 h, 6 h, 12 h, 1 d, 3 d, 1 week, 2 weeks, 3 weeks and 4 weeks after operation）. A vein thrombosis model was established by the "narrow" method. The processes of thrombosis, organization, recanalization and the features of change on hemosiderin and calcium salt were observed by HE stain, Perls stain and Von Kossa stain. The expression changes of CD61, α-SMA and CD34 were observed by immunohistochemical staining technique. RESULTS: Platelets adhered to the exposed blood vessel intima 3 h after operation, and platelet trabeculae were formed by the repeated accumulation of platelets 1 d after operation. The thrombus organization formed through the fibroblasts from vessel wall that grew into the interior of the thrombus 3 d after operation. Endothelial cells covered the surface of thrombus and then the new blood vessels were reformed, and the vessels were reconstructed. The expression of CD61 upregulated at the stages of the thrombus formation （3 h） and thrombus reformation （4 weeks）, and reached the peak 1 d after thrombus formation. The release of hemosiderin and the initial expression of α-SMA were detected 3 d later. Calcium deposit and expression of CD34 were observed 1 week later. CONCLUSIONS The hemosiderin, calcium salt, CD61, α-SMA and CD34 show time-dependent changing characteristics, which is expected to provide a reference for the estimation on thrombus formation time of the forensic cases died from thrombosis.
Animals ; Antigens, CD34/analysis* ; Hemosiderin/metabolism* ; Rats ; Venous Thrombosis/pathology*

Gastrointestinal stromal tumor (GIST) is known as considerable controversal tumor about it's histogenesis, differentiation and biologic behavior. It is traditionally regarded as smooth muscle tumor. To evaluate and clarify the origin of tumor, we performed immunohistochemical study of 23 cases of GIST on CD34 antigen, alpha-smooth muscle actin, S-100 protein, and compared the result with 4 cases of typical leiomyoma of GI tract. The results were as follows. CD34 antigen expression was noted in 21 cases (91.3%) of GIST, while typical leiomyoma was all negative. There were no difference of CD34 expression according to the biologic behavior. However, it's staining pattern was significantly different (p<0.05). Focal or multifocal expression was dominant in benign GIST (58.3%), while diffuse expression was dominant in malignant GIST (80%). Actin was expressed in 5 cases of benign GIST (38.5%) and 1 of malignant GIST (16.7%) focally. All typical leiomyoma showed diffuse strong positivity on alpha-smooth muscle actin. S-100 protein was expressed in 2 cases of benign GIST (16.7%) only. The pattern of CD34 expression was focal in the actin or S-100 protein positive cases. In conclusion CD34 antigen is useful marker in the separation of GIST, from typical smooth muscle tumor. Also it suggest that most GISTs are histogenetically primitive mesenchymal cell origin. However, CD34 expression was unrelated with biologic behavior of GIST.
Actins ; Antigens, CD34* ; Gastrointestinal Stromal Tumors* ; Gastrointestinal Tract ; Leiomyoma ; S100 Proteins ; Smooth Muscle Tumor

Spindle cell lipoma (SCL) is a rare lipoma variant that account for approximately 1.5% of all adipocyte-origin tumors; SCL usually occurs on the posterior neck or shoulder. The histological characteristics of SCL include mature, univacuolar fat cells and fibroblast-like spindle cells in a matrix of collagen and mucoid material. It is important to note that spindle cell lipoma can be mistaken both clinically and histologically for liposarcoma. We report here on a rare case of SCL in a 48-year-old male, and the patient presented with a large right neck mass that involved the lateral neck space and larynx.
Adipocytes ; Antigens, CD34 ; Collagen ; Humans ; Immunohistochemistry ; Larynx ; Lipoma ; Liposarcoma ; Male ; Middle Aged ; Neck ; Shoulder

No abstract available.
Adult ; Antigens, CD34/metabolism ; Female ; Hemangiosarcoma/diagnosis/*pathology/surgery ; Humans ; Liver Neoplasms/diagnosis/*pathology/surgery

PURPOSE: The purpose of this study was to evaluate the clinicopathological significance of the microvessel density and macrophage and mast cell counts in invasive breast carcinomas. MATERIALS AND METHODS: 45 invasive breast carcinomas were immunohistochemically stained with the endothelial antigen, CD34, and macrophage marker, CD68. 0.1% toluidine blue was used to highlight mast cells. The microvessel and mast cell counts were performed at x200 magnification and the macrophages at x400 magnification. RESULTS: With the 45 invasive breast carcinomas, there were no statistically significant associations between the mast cell, macrophage and microvessel counts and the tumor size and lymph node status. ER and PR negative mast cells infiltrated more than in cases of positive stati, with statistical significance (p-value=0.010 and 0.005, respectively). The macrophage counts were negatively correlated with the PR status (p-value=0.030). With respect to the c-erbB-2 status, there was no significance correlation with the mast cell, macrophage and microvessel counts. The mast cell counts showed significantly positive correlation with the microvessel counts in the invasive breast carcinomas (p-value=0.015). In a comparison of the macrophage counts with the microvessel counts, a positive tendency for both parameters, but without statistical significance (p-value=0.310). CONCLUSION: Increasing numbers of mast cells and macrophages were recruited in invasive breast carcinomas, which contribute to angiogenesis. The microvessel density in invasive breast carcinomas had no statistically significant association with the tumor size, lymph node status, and histological grade, presence of DCIS component, estrogen/progesterone receptor status and cerbB-2 status. The evaluation of angiogenesis using these methods is not thought to provide an independent clinicopathological factor in invasive breast carcinomas.
Antigens, CD34 ; Breast Neoplasms ; Breast* ; Carcinoma, Intraductal, Noninfiltrating ; Lymph Nodes ; Macrophages* ; Mast Cells* ; Microvessels* ; Tolonium Chloride

To investigate whether the RBC lysing solution can affect the results of relative enumeration of CD34(+) cells, 37 mobile peripheral blood apheresis products were stained using CD34-PE and CD45-FITC monoclone antibodies and RBCs were then lysed by two lysing solution commercially available (one named FACS Lysing Solution, FACS; another IOTest 3 Lysing Solution, IOTest) and one lysing solution self-prepared. After being processed by lyse-and-then-washed method, samples were detected by FACSC anto flow cytometer. The percentages of CD34(+) cells were determined based on ISHAGE gating strategy, forward and side scatter (FSC and SSC) characteristics, percentage of CD45(+) cells were recorded simultaneously. The results showed that by lyse-and-then-wash method, the percentages of CD34(+) cells in FACS-treated samples were significantly lower compared with that in IOTest-treated samples (0.50 +/- 0.42 vs 0.92 +/- 0.59, p = 0.004), but no statistical difference was observed between IOTest-treated and ourselves-prepared-treated samples. The intensities of FSC and SSC in cells of IOTest-treated sample were significantly higher compared with that in cells of FACS-treated sample (p < 0.01). The proportion of CD45(+) cells in IOTest-treated samples was lower than that in FACS-treated samples. The WBC count of samples was not correlated to the amount of CD34(+) cells (r(s) = 0.192, p = 0.357). It is concluded that the red cell lysing solution shows unexpected effect on detecting and counting CD34(+) cells, prudence should be taken to select such reagents at FCM performance.
Antigens, CD34 ; immunology ; Cell Count ; Cell Death ; Erythrocytes ; Flow Cytometry ; methods ; Humans ; Solutions ; pharmacology

OBJECTIVETo explore the rapid neutrophil engraftment and long-term hematopoietic reconstitution.

METHODSMononuclear cells (MNCs) were isolated from 5-Fu treated male BDF1 mouse bone marrow and CD(34)(+)/c-kit(+) cells were selected from the MNCs by using MoFlo Cell Sorter. MNCs and CD(34)(+)/c-kit(+) cells were co-cultured with mouse bone marrow-derived mesenchymal stem cells (MSCs) in a two-step expansion. The expanded cells were then transplanted into sublethally irradiated female BDF1 mice.

RESULTSCo-culture with MSCs resulted in 10.8-, 4.8-, 65.9- and 38.8-fold increases yields of median total nucleated cells, CD(34)(+) cells, GM-CFC and HPP-CFC, respectively, as for the MNCs culture, and 76.1-, 2.9-, 71.7- and 51.8-fold increase respectively for the CD(34)(+)/c-kit(+) cell culture. The expanded cells could rapidly engraft in the sublethally, irradiated mice, reconstitute their hematopoiesis, and be detected in the recipients bone marrow 2 months after transplantation.

CONCLUSIONSHematopoietic stem/progenitor cells co-cultures with MSCs in two-step expansion could increase expansion yields of total nucleated cells, GM-CFC and HPP-CFC. The availability of increased numbers of expanded cells may result in more rapid engraftment of neutrophils following infusion to transplant recipients.

Animals ; Antigens, CD34 ; analysis ; Hematopoiesis ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; physiology ; Male ; Mice

OBJECTIVETo investigate the mRNA expression profiles of megakaryocytes (MKs) from human cord blood CD34+ cells ex vivo expanded using Solexa technique.

METHODSCD34+ Cells were isolated using density gradient centrifugation and magnetic activated cell sorting. Cultures were stimulated with recombinant human thrombopoietin (100 ng/ml). After 12 days, the MKs fraction was separated from the non-MKs fraction using an anti-CD41 monoclonal antibody by immunomagnetic sorting. The mRNA expression of MKs and non-MKs was detected by Solexa sequencing.

RESULTSWe obtained 3 773 147 and 3 533 805 Tags from MKs and non-MKs, respectively. The amounts of unambiguous tags were 3 291 132 and 2 967 947 and those of distinct tags were 197 769 and 245 318. The expression of 1161 genes was up-regulated and that of 902 genes down-regulated. The expression of 2717 tags was up-regulated and that of 1519 tags down-regulated.

CONCLUSIONSMKs and non-MKs have remarkably different mRNA expression profiles. The differential gene-encoded products may be involved in cellular development, adhesion, apoptosis metabolism, intra- and intercellular signal transduction, and immune response. Further studies on this topic may clarify the expression mechanism, signal transduction, and regulation mechanisms.

Antigens, CD34 ; Cells, Cultured ; Fetal Blood ; cytology ; Humans ; Megakaryocytes ; cytology ; metabolism ; RNA, Messenger ; genetics ; Transcriptome