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1.
Journal of Experimental Hematology ; (6): 1807-1811, 2021.
Article in Chinese | WPRIM | ID: wpr-922339

ABSTRACT

OBJECTIVE@#To explore the diagnostic value of bone marrow cell morphology combined with immunohistochemistry in patients with primary bone marrow lymphoma.@*METHODS@#The clinical data of 23 patients with primary bone marrow lymphoma diagnosed in the First Affiliated Hospital of Xi'an Jiaotong University from January 2010 to December 2019 were collected. The characteristics of bone marrow aspiration, bone marrow biopsy and immunohistochemistry results were analyzed retrospectively, and the diagnostic value of bone marrow cell morphology combined with immunohistochemistry in primary bone marrow lymphoma were clarified.@*RESULTS@#Most of primary bone marrow lymphoma was B-cell lymphoma, among which diffuse large B-cell lymphoma was the most common pathological type. Typical lymphoma cells could be found in all the patients. 78.26% of the patients could be diagnosed as lymphoma with pathological type, while 91.30% were diagnosed as lymphoma through combined with the bone marrow immunohistochemistry.@*CONCLUSION@#Bone marrow cell morphology combined with immunohistochemistry shows very important diagnostic value in patients with primary bone marrow lymphoma.


Subject(s)
Bone Marrow , Bone Marrow Cells , Humans , Immunohistochemistry , Lymphoma, Large B-Cell, Diffuse/diagnosis , Retrospective Studies
2.
Journal of Experimental Hematology ; (6): 1631-1636, 2021.
Article in Chinese | WPRIM | ID: wpr-922307

ABSTRACT

OBJECTIVE@#To investigate the effect of high mobility group protein 1(HMGB1) on the proliferation and cytokine expression of human bone marrow mesenchymal stem cells (MSC).@*METHODS@#Different concentrations of recombinant human HMGB1 protein (100, 200, 400, 800 and 1000 ng/ml) were incubated with MSC for 24, 48, 72 h and the proliferation of MSC were detected respectively by using the CCK-8 method and flow cytometry. The best concentrations of HMGB1 incubated with MSC was determined (200 ng/ml, 1000 ng/ml), and the flow cytomerty was used to determine the effect of HMGB1 on the proliferation of MSC. The mRNA expression levels of IL-10, TGF- β1, TSG-6 and IFN-γ in MSC incubated with HMGB1 protein were detected by real-time quantitative PCR and ELISA.@*RESULTS@#The result of MSC identification and flow cytometry showed that the CD105, CD73 and CD90 were expressed, but did not expression CD45, CD34, CD11b, CD19 and HLA-DR; CCK-8 showed that HMGB1 at the concentrations of 100 ng/ml, 200 ng/ml and 400 ng/ml could promote the proliferation of MSC incubated for 24, 48 and 72 h as compared with the control group (P<0.05), and the most effective concentration was 200 ng/ml; flow cytometry showed that the compared with the control group, HMGB1 200 ng/ml could induce MSC from G1 phase to S phase to promote the proliferation of MSC; QPCR showed that the mRNA expression of MSC cytokines IL-10, TGF-β1, TSG-6 increased while IFN-γ decreased at the concentration of 200 ng/ml HMGB1 as compared with the control group. ELISA experiments showed that the HMGB1 200 ng/ml acting on MSC for 48 h could significantly promoted the secretion of IL-10, TGF-β 1 and TSG-6(P<0.05), while IFN-γ showed no significant difference as compared with control group.@*CONCLUSION@#Recombinant human HMGB1 can promote the proliferation and secretion of MSC in healthy people.


Subject(s)
Bone Marrow Cells , Cell Differentiation , Cell Proliferation , Cells, Cultured , HMGB1 Protein , Humans , Mesenchymal Stem Cells
3.
Journal of Experimental Hematology ; (6): 1224-1230, 2021.
Article in Chinese | WPRIM | ID: wpr-888542

ABSTRACT

OBJECTIVE@#To analyze the proliferation potential of bone marrow-derived mesenchymal stem cells (MSC) in patients with myelodysplastic syndrome (MDS).@*METHODS@#The MSC derived from the 24 patients with newly diagnosed MDS (MDS-MSC group) and MSC derived from 15 patients with nutritional anemia (control group) in the Affiliated Hospital of Hebei University were used as the research objects. The proliferation potential of MSC was analyzed by colony-forming unit assay, doubling time, cumulative passaging, cell number after 10 days of culture with equal amount of MSC and MTT experiment. The mechanism of abnormal proliferation was analyzed by cell cycle experiment, apoptosis experiment and p21 gene expression assay.@*RESULTS@#In the colony forming unit assay, the number of MDS-MSC colonies was 4.44±2.51, which was significantly lower than that of the control group (12.44±2.55)(P<0.01); the doubling time of MDS-MSC group was significantly longer than that of the control group (7.80±3.26 vs 3.63±0.85) (P<0.01); the number of MDS-MSC in 5×10@*CONCLUSION@#The proliferative capability of MDS-MSC is significantly reduced, which relates with the arrest of cell cycle in G


Subject(s)
Apoptosis , Bone Marrow Cells , Cell Proliferation , Humans , Mesenchymal Stem Cells , Myelodysplastic Syndromes
4.
Article in Chinese | WPRIM | ID: wpr-888337

ABSTRACT

OBJECTIVE@#To construct and identify adenovirus vector co-expressing hBMP2 and hVEGF165 fusion protein which labeled with green fluorescence protein, and laying the foundtion of the effect of hBMP2 and hVEGF165 gene inducing BMMSCs differentiation to osteoblast and bone defect repaired in the body.@*METHODS@#BMP2 and VEGF165 gene was amplified from cDNA library by PCR and inserted to the polyclonal site of adenovirus shuttle plasmid pAd-MCMV-GFP. Ad-BMP2- VEGF165 was recombinated and propagated in HEK293 cells by co-transfecting with the constructed recombinant shuttle plasmid pAd-MCMV-BMP2-VEGF165 and adenovirus helper plasmid pBHGloxΔ E1, 3Cre. The recombinant adenovirus was purified and virustiter was determined, and then to research GFP expression and to calculate the adenovirus transfection rate in rabbit BMMSCs.@*RESULTS@#The recombinant adenovirus vector Ad-BMP2-VEGF165 was successfully constructed by the methods of gene analyzing, colony PCR, Western blotting and observing GFP expression, and the titer of the adenovirus was 1×10@*CONCLUSION@#Recombinant adenovirus vector containing hBMP2 and hVEGF165 gene was successfully constructed and its high titer was obtained.


Subject(s)
Adenoviridae/genetics , Animals , Bone Marrow Cells , Genetic Vectors/genetics , HEK293 Cells , Humans , Mesenchymal Stem Cells , Rabbits , Transfection
5.
Article in English | WPRIM | ID: wpr-880681

ABSTRACT

OBJECTIVES@#To establish mouse bone marrow transplantation by pretreatment with chemotherapy, and to explore the dynamic changes of immune cells in the early stage of allogeneic transplantation in the spleen of mice.@*METHODS@#Mice were divided into 4 groups (80 mg/kg group, 100 mg/kg group, 120 mg/kg group, and 150 mg/kg group) according to the difference in dose of busulfan. The mice were treated with busulfan and cyclophosphamide combined chemotherapy, and the appropriate dosage was determined by evaluating the myeloablative effect and drug toxicity. According to the type of the genetic transplantation, the mice were also divided into 4 groups: An allogeneic transplantation group, a homogenic transplantation group, a chemotherapy alone group, and a normal control group. The mice were pretreated with busulfan and cyclophosphamide before bone marrow transplantation. In the allogeneic transplantation group, the suspension of splenocytes was prepared at the first day, the 3rd day, the 5th day, and the 8th day after transplantation for flow cytometry detection, and the dynamic changes of splenic immune cells were analyzed. The homogeneic transplantation group served as the concurrent control, the normal control group served as the control of basic value of spleen immune cells, and the chemotherapy alone group was used to evaluate the myeloablative effect.@*RESULTS@#1) The optimal dose of busulfan was 100 mg/kg. The combination of busulfan and cyclophosphamide can restore the hematopoiesis of transplanted mice, and the toxicity associated with pretreatment is small. 2) In the allogeneic transplantation group: The hematopoietic reconstitution and high donor chimerism rate were achieved after transplantation. In the early phase of bone marrow transplantation, the T lymphocytes were the main cell group, while the recovery of B lymphocytes was relatively delayed. The dendritic cells and natural killer cells from donors were the earliest cells to recover and achieve high chimerism rate compared with T cells and B cells. Most T cells were in the initial T cell state within 5 days after allogeneic transplantation. However, in the 5th day after transplantation, these cells were mainly in the effective memory phenotype. The reconstruction of donor-derived naive T cells was slow, but the reconstruction of donor-derived effective memory T cells and regulatory T cells was relatively fast. 3) In the homogeneic transplantation group: The mice could recover hematopoiesis and the recovery of B lymphocytes was delayed. 4) In the chemotherapy alone group: All mice died in 12-15 days after chemotherapy, and the peripheral blood routine showed pancytopenia before death.@*CONCLUSIONS@#Pretreatment with chemotherapy can successfully establish the mouse model of bone marrow transplantation. There are difference in the proportion of T cells, B cells, natural killer cells, dendritic cells, effector memory T cells, initial T cells, and regulatory T cells after transplantation, and the relationship between donor and recipient is also changed.


Subject(s)
Animals , Bone Marrow Cells , Bone Marrow Transplantation , Busulfan , Cell Proliferation , Kinetics , Mice , Mice, Inbred C57BL , Transplantation, Homologous
6.
Article in English | WPRIM | ID: wpr-880667

ABSTRACT

OBJECTIVES@#Excessive production of AGEs in diabetic patients will affect the normal function of osteoblasts, and this process may be related to autophagy of osteoblasts. This study aims to explore the effect of advanced glycation end products (AGEs) on autophagic activity during osteogenic differentiation in rat bone marrow mesenchymal stem cells (BMSCs).@*METHODS@#BMSCs were isolated and cultured in vitro, treated with different concentrations (0, 50, 100, 200, and 400 mg/L) of AGEs for different time (3, 6, 12, 24, 48, and 72 h). The proliferation activity was detected by CCK-8 method. The mRNA and protein expression levels of Beclin1 and LC3 in cells were detected by real-time PCR and Western blotting, respectively.The autophagic vacuoles were observed under the transmission electron microscope. The cells were treated with autophagy promoter rapamycin or autophagy inhibitor 3MA. After 7 days of osteogenic induction, we performed alkaline phosphatase (ALP) staining and real-time PCR to detect the mRNA expression levels of osteogenesis-related genes.@*RESULTS@#In the low-concentration groups, the proliferation activity in BMSCs was increased (@*CONCLUSIONS@#Low concentration of AGEs can enhance the proliferative activity of BMSCs and promote osteogenic differentiation by accelerating autophagy. High concentration of AGEs can suppress the proliferation of BMSCs and inhibit osteogenic differentiation by reducing autophagy.


Subject(s)
Animals , Autophagy , Bone Marrow Cells , Cell Differentiation , Cells, Cultured , Glycation End Products, Advanced/pharmacology , Humans , Osteoblasts , Osteogenesis , Rats , Rats, Sprague-Dawley
7.
Journal of Experimental Hematology ; (6): 1002-1006, 2021.
Article in Chinese | WPRIM | ID: wpr-880183

ABSTRACT

Emerging data have demonstrated that bone marrow mesenchymal stem cells (MSCs) play important roles in the progression of myelodysplastic syndrome (MDS). Experiments in vitro have showed that MSCs derived from MDS patients (MDS-MSC) exhibit the biological characteristics of cell senescence. Although the underlying mechanisms that regulate cell senescence need to be further elucidated, existing researches indicate that the mechanisms of MDS-MSC senescence have significant heterogeneity. Depth understanding of the underlying mechanisms involved in cell senescence of MDS-MSC are crucial to explore the potential therapeutic target of MDS. Therefore, this review summarizes research advances related with MSC senescence, such as MDS-MSC intrinsic changes in telomere shortening, DNA methylation status, oxidative stress and signal pathways regulating cell senescence in recent years.


Subject(s)
Bone Marrow , Bone Marrow Cells , Cellular Senescence , Humans , Mesenchymal Stem Cells , Myelodysplastic Syndromes
8.
Article in Chinese | WPRIM | ID: wpr-880155

ABSTRACT

OBJECTIVE@#To investigate the quantitative expression of immunophenotype of CD34@*METHODS@#Multi-parameter flow cytometry (FCM) was used to detect the proportion and mean fluorescence intensity (MFI) of each antigen of bone marrow CD34@*RESULTS@#Bone marrow blast cell proportion (P<0.01), RBC level (P<0.01), and Hb level (P<0.05) of high-risk MDS patients were higher, while EPO level (P<0.05) was lower than those of low-risk patients. The proportion of CD34@*CONCLUSION@#The immunophenotype of CD34


Subject(s)
Antigens, CD34 , Bone Marrow , Bone Marrow Cells , Flow Cytometry , Humans , Immunophenotyping , Myelodysplastic Syndromes
9.
Article in Chinese | WPRIM | ID: wpr-880113

ABSTRACT

METHODS@#To establish the acquired aplastic anemia mouse model through the X-ray irradiation in combination with lymphocytes injection. AA Group: the purified Pan T lymphocytes from the spleen of C57BL/6J mice were enriched and injected to the mice through tail vein(5×10@*RESULTS@#Compared with 4, 5 Gy irradiated mice in AA groups, the survival time of 3 Gy irradiated AA groups was significantly prolonged. 3, 4 and 5 Gy X-ray irradiation combined with Pan T lymphocyte injection could successfully induced severe reduction of red blood cells, blood neutrophils, and platelets, severe reduction of bone marrow nucleated cells, severe bone marrow hematopoietic failure, and the significant expansion of T lymphocytes ratio in the bone marrow. CD4@*CONCLUSION@#3, 4 and 5 Gy X-ray irradiation combined with 5×10


Subject(s)
Anemia, Aplastic , Animals , Bone Marrow , Bone Marrow Cells , CD8-Positive T-Lymphocytes , Humans , Mice , Mice, Inbred C57BL
10.
Article in Chinese | WPRIM | ID: wpr-880108

ABSTRACT

OBJECTIVE@#To analyze the influence of serum homocysteine (Hcy) levels to the prognosis of newly diagnosed multiple myeloma (MM) patients, and to explore related factors affecting the prognosis of the patients.@*METHODS@#The clinical pathological data of 180 newly diagnosed MM patients treated in our hospital from March 2013 to February 2015 were collected, and the patients were divided into high and low Hcy groups based on the median Hcy. The survival curves of the patients in the two groups were drawn to compare the differences of the survival; univariate and multivariate survival analysis was used to observe the influence of serum cysteine to the prognosis of newly diagnosed MM patients; the clinicopathological data of the patients with high and low Hcy in the two groups was compared, Pearson test was used to further analyzes the relationship between Hcy and different factors, and explores the related factors of Hcy affecting the prognosis of the patients.@*RESULTS@#The median survival times of patients in the high and low Hcy groups were 32 (5-59) and 41 (7-71) months, respectively. The 3-year survival rate of the patients in high Hcy group was significantly lower than those in low Hcy group, and the difference shows statistically significant (P<0.05). The results of univariate survival analysis showed that the OS of newly diagnosed MM patients whom with advanced age, high bone disease grade, high-level bone marrow plasma cell count, LDH, C-reactive protein, Cr, β@*CONCLUSION@#Serum Hcy level has a correlation trend with the survival of newly diagnosed MM, which is affected by factors such as Hb.


Subject(s)
Bone Marrow Cells , Homocysteine , Humans , Multiple Myeloma , Prognosis , Risk Factors
11.
Article in Chinese | WPRIM | ID: wpr-880072

ABSTRACT

The normal hematopoiesis of the body are depends on the proliferation and differentiation of hematopoietic stem/progenitor cell (HSPC), as well as the mesenchymal stem/stromal cell (MSC) that support the growth and development of hematopoietic microenvironment of bone marrow (BM). At present, the commonly used MSC for promoting hematopoiesis are mainly derived from BM, however, the acquisition of MSC from BM is limited by the number, sampling, isolation and purification. Compared with BM, adipose tissue has many advantages, including widely distributed, abundant in source, simple and easy to obtain, and contains more pluripotent stem cell, such as adipose tissue-derived stem cell (ADSC), in which Perivascular cell/pericyte (PC) are considered as the precursor cell of MSC, also is the main components of vascular microenvironment, and plays an important role in the proliferation and differentiation of HSPC. PC is especially abundant in adipose tissue, and with the advantages of easy acquisition, small damage, fast cell proliferation and low immunogenicity. Therefore, the sustaining hematopoiess of human adipose derived-perivascular stem cell (AD-PC) to HSPC requires further research and exploration. In this review, the possible supporting effects and PC subgroup of ADSC as stromal cell on HSPC are summarized briefly.


Subject(s)
Adipose Tissue , Bone Marrow Cells , Cell Differentiation , Cell Proliferation , Cells, Cultured , Hematopoietic Stem Cell Transplantation , Hematopoietic Stem Cells , Humans , Mesenchymal Stem Cells
12.
Rev. med. Risaralda ; 26(2): 154-156, jul.-dic. 2020. graf
Article in Spanish | LILACS, COLNAL | ID: biblio-1150023

ABSTRACT

Resumen Introducción: Los linfomas de células T son infrecuentes y se caracterizan por presentarse en la población de los adultos jóvenes. Además, suele acompañarse de patologías como la anemia moderada, hepatoesplenomegalia y trombocitopenia e infiltración sinusoidal por linfocitos T en células de médula ósea, bazo e hígado. Caso clínico: Se presenta un caso clínico de un adolescente que tiene los síntomas característicos de esta patología, con sospecha clínica y diagnóstico paraclínico confirmado con histoquímica de médula ósea. Conclusión: Es una entidad infrecuente de pronóstico desfavorable, hasta el momento el paciente está estable recibiendo tratamiento. Para utilizar el enfoque adecuado en el diagnóstico y brindar tratamiento, es necesario considerar todos los hallazgos clínicos.


Abstract Introduction: T-cell lymphomas are uncommon; these tend to be present in young adult patients. Additionally, this condition is characterized by the existence of pathologies like moderate anemia, hepatosplenomegaly disorder, thrombocytopenia and sinusoidal infiltration by T-Lymphocytes in bone marrow cells, spleen and liver. In this study a case of this rare lymphoma is going to be presented. Case report: A clinical case of an adolescent who presents the characteristic symptoms of this pathology is exposed. This clinical suspicion held a paraclinical diagnosis that was confirmed by histochemistry of bone marrow tests. Conclusion: It is an infrequent condition with an unfavorable prognosis. Until now the patient remains stable and is receiving treatment, the clinical findings of the disease raise awareness about the importance of carrying out the appropriate diagnosis procedures and providing treatment.


Subject(s)
Humans , Adolescent , Splenomegaly , Thrombocytopenia , Bone Marrow Cells , T-Lymphocytes , Lymphoma, T-Cell , Hepatomegaly , Spleen , Therapeutics , Bone Marrow , Anemia , Liver
14.
Acta Physiologica Sinica ; (6): 597-604, 2020.
Article in Chinese | WPRIM | ID: wpr-878205

ABSTRACT

Pulmonary fibrosis is a chronic, diffuse, interstitial lung disease involving the pulmonary interstitium, alveoli, and bronchioles caused by various causes. There is no effective treatment. Currently, exogenous bone marrow-derived mesenchymal stem cells (BM-MSCs) transplantation has attracted much attention as a new stem cell therapy in the treatment of pulmonary fibrosis. Less attention has been paid to the functional status of endogenous BM-MSCs during pulmonary fibrosis. Based on summary on the anti-pulmonary fibrosis effect of BM-MSCs and its mechanism, this review further discusses the abnormal changes of bone marrow function in animals with pulmonary fibrosis and the role of glutamate NMDA receptor overactivation in mediating the functional inhibition of endogenous BM-MSCs induced by pulmonary fibrosis. This will provide potential ideas for finding effective treatments for pulmonary fibrosis.


Subject(s)
Animals , Bone Marrow , Bone Marrow Cells , Mesenchymal Stem Cells , Pulmonary Fibrosis , Signal Transduction
15.
Article in Chinese | WPRIM | ID: wpr-827184

ABSTRACT

OBJECTIVE@#To establish a stable and rapidly rat model acquired aplastic anemia.@*METHODS@#The SD rats were exposed to Csγ-ray at 3.5 and 4.0 Gy ( 91 cGy/min), and intraperitoneally injected with CTX at 35 mg/( kg·d) and CHL at 45 mg/( kg·d) in d 4, 6 and 8 after irradiation; the WBC, platelet and reticulocyte counts in peripheral blood, the smears and nucleated cells counts of bone marrow were observed.@*RESULTS@#The levels of peripheral blood 3-lineage cells of SD rats treated with Csγ-irradiation combined with cyclophosphamide and chloramphenicol were significantly reduced, among which white blood cells, platelets and reticulocytes decreased rapidly, and the number of bone marrow nucleated cells decreased significantly; bone marrow pathological sections showed severe reduction of hematopoietic cells, and the non-hematopoietic cells such as fat cells increased, and a serve or lightly reduction of bone marrow cells were found.@*CONCLUSION@#The rat model established by Csγ-ray irradiation combined with cyclophosphamide and chloramphenicol meets the clinical characteristics of aplastic anemia, and this study provides a stable rat model for the study of new therapeutic drugs for acquired aplastic anemia.


Subject(s)
Anemia, Aplastic , Animals , Bone Marrow , Bone Marrow Cells , Cyclophosphamide , Rats , Rats, Sprague-Dawley
16.
Journal of Experimental Hematology ; (6): 1032-1037, 2020.
Article in Chinese | WPRIM | ID: wpr-827166

ABSTRACT

OBJECTIVE@#To explore the effect of mmu-circRNA_016901 on the regulation of radiation injury of bone marrow stem cells and its mechanism.@*METHODS@#Bone marrow stem cells were exposed to different dose of X-ray, then the expression level of mmu-circRNA_016901 in bone marrow cells treated with different doses of X-ray was detected. The luciferase reporter gene assay was used to confirm that miRNA1249-5p is the target of mmu-circRNA_016901, and RNA Binding Protein Immunoprecipitation was used to confirm that TGF-β3 is the targeted on miRNA1249-5p,the expression of TGF-β3 and cell proliferation were detected after the expression of mmu-circRNA_01690 was regulated.@*RESULTS@#When the irradiation dose<6 Gy, there were significant difference in the expression of mmn-circRNA-016901 after the bone marrow mesenchymal stem cells were treated by different doses of irradiation, which showed a statistically significant (P<0.05). The luciferase reporter gene detection and co-immunoprecipitation experiments confirmed that Mmu-circRNA_016901 could binds to miRNA1249-5p specifically, and overexpression of mmu-circRNA_016901 could regulate miRNA1249-5p negatively, which resulted in a significant increase in TGF-β3 expression and promoting of cell proliferation.@*CONCLUSION@#mmu-circRNA_016901 affects the expression of TGF-β3 through miRNA1249-5p, and thus participates in the regulation of the radiation damage mechanism of bone marrow mesenchymal stem cells.


Subject(s)
Bone Marrow Cells , Mesenchymal Stem Cells , RNA, Circular , Genetics , Radiation Tolerance
17.
Journal of Experimental Hematology ; (6): 1157-1161, 2020.
Article in Chinese | WPRIM | ID: wpr-827147

ABSTRACT

OBJECTIVE@#To study the expression of Peroxiredoxin-6 (Prdx6) gene in elderly patients with acute myeloid leukemia (AML) and its clinical significance.@*METHODS@#The expression level of Prdx6 in bone marrow cells of 33 cases of AML, 8 cases of CML and 11 cases of other blood diseases was detected by PCR. The correlation of the abnormal expression of Prdx6 with patient age and blood routine parameters was further analyzed.@*RESULTS@#the expression level of Prdx6 in elderly patients with AML (≥60 years) was significantly lower than that in younger patients (<60 years) (P<0.05); the expression level of Prdx6 in low WBC (≤1×10/L) group was lower than that in medium WBC (1-10×10/L) group or high WBC (>10×10/L) group (P<0.05); the proportion of WBC count (≤1×10/L) in elderly patients with AML reached 38.5%, which was significantly higher than that in younger patients (5%) (P<0.05); the overall survival (OS) rate of elderly patients was lower than that of younger patients (P<0.05).@*CONCLUSION@#The expression of Prdx6 in elderly patients with AML is low, moreover, it relates with low value of WBC in peripheral blood, suggesting that it may be one of poor prognostic factors for the elderly patients with AML.


Subject(s)
Aged , Bone Marrow Cells , Humans , Leukemia, Myeloid, Acute , Leukocyte Count , Peroxiredoxin VI , Genetics , Metabolism , Prognosis
18.
Journal of Experimental Hematology ; (6): 1349-1356, 2020.
Article in Chinese | WPRIM | ID: wpr-827113

ABSTRACT

OBJECTIVE@#To investigate the effect of PDGFRα stromal cells derived SCF on hematopoiesis of adult mice.@*METHODS@#Pdgfrα-CreER; R26-tdTomato mice model was constructed, and the proportion and distribution of PDGFRα cells in the liver, spleen, lung, kidney and bone marrow were analyzed by flow cytometry and confocal microscopy. Then the Pdgfrα-CreER; Scf mice model was further constructed, the Scf in PDGFRα was knocked out specifically, the effect of Scf-knocked out in PDGFRα stromal cells in the propitiation of HSPCs in the bone marrow was analyzed by flow cytometry. The effect of SCF on the proportion on number of peripheral blood cells in mice was analyzed by whole blood analyzer.@*RESULTS@#After Scf was knocked out in PDGFRα stromal cells, the propitiation and number of LKS- cell, LKS+ cell, HSC, MPP1, MKP, PreGM, PreMegE, and CFU-E in the bone marrow of mice was decreased, as well as in the number of red blood cells and hemoglobin concentration of peripheral blood. However, Scf knocked out from PDGFRα cells showed no effect on the hematopoiesis in spleen.@*CONCLUSION@#specific knocked out of Scf in PDGFRα stromal cells in adult mice can decrease the proportion of HSPCs in the bone marrow and the number of red blood cells in peripheral blood, and finally lead to anemia in mice.


Subject(s)
Animals , Bone Marrow , Bone Marrow Cells , Hematopoiesis , Mice , Receptor, Platelet-Derived Growth Factor alpha , Stem Cell Factor
19.
Journal of Experimental Hematology ; (6): 1357-1362, 2020.
Article in Chinese | WPRIM | ID: wpr-827112

ABSTRACT

OBJECTIVE@#To explore the method for inducing the differentiation of bone marrow cells into megakaryocytes in vitro so as to use for evaluating the activity of traditional Chinese medicines.@*METHODS@#The bone marrow cells were separated from femurs and tibias of mice. The experiments were divided into 4 groups: control (no adding cytokines), TPO (adding 50 ng/ml TPO), TPO+SCF (50 ng/ml+50 ng/ml) and TPO+SCF+IL-6+IL-9 (50 ng/ml+50 ng/ml+20 ng/ml+20 ng/ml). The bone marrow cells in 4 groups were cultured in vitro for 6 d. Then the cell growth status was observed by the inverted microscopy, and the cell count was detected by using the automatic cell counter. The ratio and absolute count of megakaryocytes were detected by flow cytometry.@*RESULTS@#Compared with control, three induction methods could stimulate the differentiation of bone marrow cells into megakaryocytes in vitro. TPO could slightly enhance the differentiation of bone marrow cells into megakaryocytes. Both the combination of TPO and SCF, and the combination of TPO, SCF, IL-6 and IL-9 could intensively stimulate proliferation of bone morrow cells and promote the differentiation of bone marrow cells into megakaryocytes. The addition of IL-6 and IL-9 could decrease the proliferation of non-megakaryocytes, but promote the differentiation of bone marrow cells into megakaryocytes.@*CONCLUSION@#The optimized differentiation of bone marrow cells into megakaryocytes has been completed by co-induction regimen of TPO, SCF, IL-6 and IL-9, which can be used to screen and evaluate traditional Chinese medicines promoting formation of platelets.


Subject(s)
Animals , Bone Marrow Cells , Cell Count , Cell Differentiation , Cell Division , Cells, Cultured , Interleukin-3 , Megakaryocytes , Mice , Stem Cell Factor , Thrombopoietin
20.
Acta Physiologica Sinica ; (6): 167-174, 2020.
Article in Chinese | WPRIM | ID: wpr-827071

ABSTRACT

Humans with chronic psychological stress are prone to develop multiple disorders of body function including impairment of immune system. Chronic psychological stress has been reported to have negative effects on body immune system. However, the underlying mechanisms have not been clearly demonstrated. All immune cells are derived from hematopoietic stem cells (HSC) in the bone marrow, including myeloid cells which comprise the innate immunity as a pivotal component. In this study, to explore the effects of chronic psychological stress on HSC and myeloid cells, different repeated restraint sessions were applied, including long-term mild restraint in which mice were individually subjected to a 2 h restraint session twice daily (morning and afternoon/between 9:00 and 17:00) for 4 weeks, and short-term vigorous restraint in which mice were individually subjected to a 16 h restraint session (from 17:00 to 9:00 next day) for 5 days. At the end of restraint, mice were sacrificed and the total cell numbers in the bone marrow and peripheral blood were measured by cell counting. The proportions and absolute numbers of HSC (LinCD117Sca1CD150CD48) and myeloid cells (CD11bLy6C) were detected by fluorescence activated cell sorting (FACS) analysis. Proliferation of HSC was measured by BrdU incorporation assay. The results indicated that the absolute number of HSC was increased upon long-term mild restraint, but was decreased upon short-term vigorous restraint with impaired proliferation. Both long-term mild restraint and short-term vigorous restraint led to the accumulation of CD11bLy6C cells in the bone marrow as well as in the peripheral blood, as indicated by the absolute cell numbers. Taken together, long-term chronic stress led to increased ratio and absolute number of HSC in mice, while short-term stress had opposite effects, which suggests that stress-induced accumulation of CD11bLy6C myeloid cells might not result from increased number of HSC.


Subject(s)
Animals , Antigens, Ly , Metabolism , Bone Marrow Cells , Cell Biology , CD11b Antigen , Metabolism , Cell Proliferation , Hematopoietic Stem Cells , Cell Biology , Mice , Mice, Inbred C57BL , Restraint, Physical , Stress, Psychological
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