Research progress in lineage tracing to explore hepatic parenchymal cell regeneration and repair mechanisms / 中华肝脏病杂志

Hepatic parenchymal cells are a type of liver cells that performs important functions such as metabolism and detoxification. The contribution of hepatic parenchymal cells, bile duct cells, and hepatic stem/progenitor cells to new hepatic parenchymal cells in the process of liver injury repair has become a controversial issue due to their strong proliferation ability. Lineage tracing technology, which has emerged in the past decade as a new method for exploring the origin of cells, can trace specific type of cells and their daughter cells by labeling cells that express the specific gene and their progeny. The article reviews the current literature on the origin and contribution of hepatic parenchymal cells by this technique. About 98% of new hepatic parenchymal cells originate from the existing hepatic parenchymal cells during liver homeostasis and after acute injury. However, under conditions of severe liver injury, such as inhibition of hepatic parenchymal cell proliferation, bile duct cells (mainly liver stem/progenitor cells) become the predominant source of hepatic parenchymal cells, contributing a steady increased hepatocyte regeneration with the extension of time.

Differentiation of human AB plasma hematopoietic stem/progenitor cells from peripheral blood into mature erythrocytes / 中国输血杂志

【Objective】 To investigate the feasibility of differentiation of human AB plasma hematopoietic stem/progenitor cells (HSCs/HPCs) from peripheral blood into mature erythrocytes. 【Methods】 Hematopoietic stem/progenitor cells were induced to be differentiated into mature erythrocytes in the medium supplemented with 5% FBS, 3% FBS + 2% human AB plasma and 8% human AB plasma, respectively, and inoculated in 24-well culture plate at the density of 1×106/mL. Cell proliferation and morphological changes were observed in three different groups. Flow cytometry was used to detect erythroid terminal differentiation markers, i. e. GPA, Band3 and α4(α4-integrin), and late erythroid cell enucleation in different group. The effects of different culture conditions on HSCs/HPCs differentiation into mature erythrocytes were compared. 【Results】 The cell growth and proliferation multiples of the three groups (8% human AB plasma, 5% FBS and 3% FBS+ 2% human AB plasma) were 2 573±116 vs 2 514±246 vs 2 539±119(P>0.05), respectively. The morphological changes of the three groups were similar. With the extension of culture time, the cells differentiated from proerythroblasts to basophils, polychromatic erythroblasts and positive erythroblasts, and almost all of them differentiated into erythrocytes enucleation on day 21. GPA expression and enucleation rate(%) of the three groups were 97.17±1.91 vs 94.95±1.61 vs 96.15±1.38, and 85.1±3.26 vs 86.93±5.96 vs 86.5±3.36(P>0.05), respectively. 【Conclusion】 The differentiation of HSCs/HPCs from peripheral blood plasma into mature erythrocytes from human AB was similar to that of fetal bovine serum.

Biological characteristics of chondrocytes derived from auricles of different ages / 中华整形外科杂志

Objective@#To investigate the tissue structure, chondrocyte characteristics, and the differential expression of related genes and cell surface markers of auricular cartilage of patients in different ages, in order to provide a basis for the age selection of tissue engineered cartilage repair defects.@*Methods@#The auricular cartilage tissue was obtained from 22 patients with microtia in the Plastic Surgery Hospital, Chinese Academy of Medical Science, ranged from 6 to 28 years old, and divided into the child group (6-12 years old), the adolescent group (13-18 years old) and the adult group (21-28 years old). The proliferation and differentiation features of chondrocytes which from different-aged patients were detected. Furthermore, quantitative real-time PCR was used to detect the differences in the expression of genes related to cell proliferation and chondrocyte extracellular matrix. Flow cytometry, immunofluorescence and immunohistochemistry were used to detect the differences in the expression of mesenchymal stem cell markers CD90, CD44, CD73 and CD105 in chondrocytes. SPSS Statistics 21.0 software was used to process statistics.@*Results@#The proliferation capability of auricular chondrocytes of children was stronger than adolescents and adults, the child group vs the adult group P<0.05, the child group vs the adolescent group P<0.01. The expression of cartilage extracellular matrix related gene COL2A1 increased with age, the child group vs the adult group P<0.01, the adolescent group vs the adult group P<0.01. While the capability of cell osteogenic differentiation decreased with age(P<0.05). However, there was no significant difference in the capability of adipogenic differentiation when considering the ages of patients. The results of both flow cytometry and real-time PCR showed that the expression of mesenchymal stem cell markers decreased with age, with the most significant decrease in CD90(P<0.01).@*Conclusions@#The biological characteristics and stem cell content of cells derived from auricular cartilage tissue was influenced by the patients′age.

Glial Cell Line-derived Neurotrophic Factor-overexpressing Human Neural Stem/Progenitor Cells Enhance Therapeutic Efficiency in Rat with Traumatic Spinal Cord Injury

Spinal cord injury (SCI) causes axonal damage and demyelination, neural cell death, and comprehensive tissue loss, resulting in devastating neurological dysfunction. Neural stem/progenitor cell (NSPCs) transplantation provides therapeutic benefits for neural repair in SCI, and glial cell line-derived neurotrophic factor (GDNF) has been uncovered to have capability of stimulating axonal regeneration and remyelination after SCI. In this study, to evaluate whether GDNF would augment therapeutic effects of NSPCs for SCI, GDNF-encoding or mock adenoviral vector-transduced human NSPCs (GDNF-or Mock-hNSPCs) were transplanted into the injured thoracic spinal cords of rats at 7 days after SCI. Grafted GDNF-hNSPCs showed robust engraftment, long-term survival, an extensive distribution, and increased differentiation into neurons and oligodendroglial cells. Compared with Mock-hNSPC- and vehicle-injected groups, transplantation of GDNF-hNSPCs significantly reduced lesion volume and glial scar formation, promoted neurite outgrowth, axonal regeneration and myelination, increased Schwann cell migration that contributed to the myelin repair, and improved locomotor recovery. In addition, tract tracing demonstrated that transplantation of GDNF-hNSPCs reduced significantly axonal dieback of the dorsal corticospinal tract (dCST), and increased the levels of dCST collaterals, propriospinal neurons (PSNs), and contacts between dCST collaterals and PSNs in the cervical enlargement over that of the controls. Finally grafted GDNF-hNSPCs substantially reversed the increased expression of voltage-gated sodium channels and neuropeptide Y, and elevated expression of GABA in the injured spinal cord, which are involved in the attenuation of neuropathic pain after SCI. These findings suggest that implantation of GDNF-hNSPCs enhances therapeutic efficiency of hNSPCs-based cell therapy for SCI.

The current application of stem cell based therapy in pulmonary hypertension / 药学学报

Pulmonary hypertension (PH) is a progressive cardiopulmonary disease with high mortality and increased prevalence. The target therapy of pulmonary hypertension is mainly dependent on vasodilation, but how to improve vascular remodeling with stem cell therapy has not been fully understood. Over the past ten years, the researches on endothelial progenitor cells, mesenchymal stem cells and pluripotent cells have brought the hope to patients with pulmonary hypertension. This article mainly introduces the latest progress of stem cell technology in the intervention of pulmonary hypertension, and discusses the problems in the application.

Curcumin Increase the Expression of Neural Stem/Progenitor Cells and Improves Functional Recovery after Spinal Cord Injury

OBJECTIVE: To investigates the effect of curcumin on proliferation of spinal cord neural stem/progenitor cells (SC-NSPCs) and functional outcome in a rat spinal cord injury (SCI) model. METHODS: Sixty adult male Sprague-Dawley rats were randomly and blindly allocated into three groups (sham control group; curcumin treated group after SCI; vehicle treated group after SCI). Functional recovery was evaluated by the Basso, Beattie, and Bresnahan (BBB) scale during 6 weeks after SCI. The expression of SC-NSPC proliferation and astrogliosis were analyzed by nestin/Bromodeoxyuridine (BrdU) and Glial fibrillary acidic protein (GFAP) staining. The injured spinal cord was then examined histologically, including quantification of cavitation. RESULTS: The BBB score of the SCI-curcumin group was better than that of SCI-vehicle group up to 14 days (p < 0.05). The co-immunoreactivity of nestin/BrdU in the SCI-curcumin group was much higher than that of the SCI-vehicle group 1 week after surgery (p < 0.05). The GFAP immunoreactivity of the SCI-curcumin group was remarkably lower than that of the SCI-vehicle group 4 weeks after surgery (p < 0.05). The lesion cavity was significantly reduced in the curcumin group as compared to the control group (p < 0.05). CONCLUSION: These results indicate that curcumin could increase the expression of SC-NSPCs, and reduce the activity of reactive astrogliosis and lesion cavity. Consequently curcumin could improve the functional recovery after SCI via SC-NSPC properties.

Curcumin Increase the Expression of Neural Stem/Progenitor Cells and Improves Functional Recovery after Spinal Cord Injury

OBJECTIVE: To investigates the effect of curcumin on proliferation of spinal cord neural stem/progenitor cells (SC-NSPCs) and functional outcome in a rat spinal cord injury (SCI) model.METHODS: Sixty adult male Sprague-Dawley rats were randomly and blindly allocated into three groups (sham control group; curcumin treated group after SCI; vehicle treated group after SCI). Functional recovery was evaluated by the Basso, Beattie, and Bresnahan (BBB) scale during 6 weeks after SCI. The expression of SC-NSPC proliferation and astrogliosis were analyzed by nestin/Bromodeoxyuridine (BrdU) and Glial fibrillary acidic protein (GFAP) staining. The injured spinal cord was then examined histologically, including quantification of cavitation.RESULTS: The BBB score of the SCI-curcumin group was better than that of SCI-vehicle group up to 14 days (p < 0.05). The co-immunoreactivity of nestin/BrdU in the SCI-curcumin group was much higher than that of the SCI-vehicle group 1 week after surgery (p < 0.05). The GFAP immunoreactivity of the SCI-curcumin group was remarkably lower than that of the SCI-vehicle group 4 weeks after surgery (p < 0.05). The lesion cavity was significantly reduced in the curcumin group as compared to the control group (p < 0.05).CONCLUSION: These results indicate that curcumin could increase the expression of SC-NSPCs, and reduce the activity of reactive astrogliosis and lesion cavity. Consequently curcumin could improve the functional recovery after SCI via SC-NSPC properties.

Effect of oxygen glucose deprivation-reperfusion in astrocytes overexpressing endothelin-1 on the proliferation of neural stem/progenitor cells / 器官移植

Objective To investigate the effect of oxygen glucose deprivation-reperfusion (OGD-R) in astrocytes overexpressing endothelin (ET)-1 on the proliferation of neural stem/progenitor cells (NSPCs). Methods OGD-R models of negative control astrocytes (C6-Mock) and astrocytes over-expressing ET-1 (C6-ET-1) were constructed. Transwell co-culture system of astrocytes and NSPCs was established. Morphologic observation and identification of the astrocytes and primary NSPCs were performed. The cells were divided into four groups: C6-Mock+NSPCs, OGD-R+C6-Mock+NSPCs, C6-ET-1+NSPCs and OGD-R+C6-ET-1+NSPCs groups and co-cultured for 0, 24, 48 and 72 h respectively. The diameter of neurosphere was measured in each group. Results In the C6-Mock and C6-ET-1 cells, type Ⅰ astrocytes in fibrous morphology were observed. Glial fibrillary acidic protein (GFAP) was expressed in the cytoplasm of these two types of cells. Primary NSPCs were positive for nestin staining. After co-culture for 48 and 72 h, the neurosphere diameter in the OGD-R+C6-Mock+NSPCs group was significantly greater than that in the C6-Mock+NSPCs group. The neurosphere diameter in the OGD-R+C6-ET-1+NSPCs group was considerably greater than that in the C6-ET-1+NSPCs group. The neurosphere diameter in the OGD-R+C6-ET-1+NSPCs group was significantly greater compared with that in the OGD-R+C6-Mock+NSPCs group (all P<0.05). Conclusions OGD-R astrocytes can promote the proliferation of NSPCs. ET-1 over-expression further accelerates the proliferation of NSPCs.

Effect of ginsenoside Rg1 on senescence characteristics of leukemic stem cells / 局解手术学杂志

Objective To explore the effect of ginsenoside Rg1 on leukemia stem cells through comparing the biological senescence characteristics of HSCs in the patients with leukemia and healthy people,and provide new ideas and methods for leukemia prevention and treatment.Methods Fifteen cases of normal bone marrow in normal group and sixteen cases of chronic myeloid leukemia in leukemia group were divided into control group and Rg1 group,respectively.The control group used the conventional culture.The Rg1 group used the culture system with 10 μg/mL ginsenoside Rg1,other conditions were the same as control group.The bone marrow mononuclear cell of all groups were extracted after 2 days,and the CD34 +/CD38-cells population was isolated and purified by immunomagnetic adsorption cell sorting(MACS).The purity of the cells and cell cycles phase were detected by flow cytometry.Cell viability was detected by trypan blue staining.The percentage of positive cells was detected by SA-β-gal staining.CCK-8 detected the CD34 +/CD38-proliferation ability of each group.Results The ratio of CD34 +/CD38-cell population was (1.76 ± 0.34) ％ in every 1 × 106 BMNCs before sorting;the proportion of CD34 +/CD38-cell population per 1 × 106 cells after immunomagnetic sorting was (91.15 ± 2.41)％.The positive rate of SA-β-gal staining in human bone marrow CD34 +/CD38-cells of leukemia Rg1 group was significantly higher than that in leukemia control group,the difference was not significant (P ＞ 0.05);meanwhile there was no significant difference between normal control group and normal Rg1 group,but higher than that in leukemia control group,the difference was significant(P ＜ 0.05).CCK-8 results showed that the proliferation of CD34 +/CD38-cells was significantly increased in leukemia control group than those in the other groups.The survival rate of CD34 +/CD38-cells in human bone marrow was 99.1％ in all groups.Cell cycle phase results showed that the G1 arrest of CD34 +/CD38-cells in leukemia control group was significantly lower than those in the other three groups.Conclusion CD34 +/CD38-cells in chronic myeloid leukemia patients may be caused by some chronic myeloid leukemia.Ginsenoside Rg1 can effectively delay the process of aging.

Generation of Induced Pluripotent Stem Cells and Neural Stem/Progenitor Cells from Newborns with Spina Bifida Aperta

STUDY DESIGN: We established induced pluripotent stem cells (iPSCs) and neural stem/progenitor cells (NSPCs) from three newborns with spina bifida aperta (SBa) using clinically practical methods. PURPOSE: We aimed to develop stem cell lines derived from newborns with SBa for future therapeutic use. OVERVIEW OF LITERATURE: SBa is a common congenital spinal cord abnormality that causes defects in neurological and urological functions. Stem cell transplantation therapies are predicted to provide beneficial effects for patients with SBa. However, the availability of appropriate cell sources is inadequate for clinical use because of their limited accessibility and expandability, as well as ethical issues. METHODS: Fibroblast cultures were established from small fragments of skin obtained from newborns with SBa during SBa repair surgery. The cultured cells were transfected with episomal plasmid vectors encoding reprogramming factors necessary for generating iPSCs. These cells were then differentiated into NSPCs by chemical compound treatment, and NSPCs were expanded using neurosphere technology. RESULTS: We successfully generated iPSC lines from the neonatal dermal fibroblasts of three newborns with SBa. We confirmed that these lines exhibited the characteristics of human pluripotent stem cells. We successfully generated NSPCs from all SBa newborn-derived iPSCs with a combination of neural induction and neurosphere technology. CONCLUSIONS: We successfully generated iPSCs and iPSC-NSPCs from surgical samples obtained from newborns with SBa with the goal of future clinical use in patients with SBa.

M2 Phenotype Microglia-derived Cytokine Stimulates Proliferation and Neuronal Differentiation of Endogenous Stem Cells in Ischemic Brain

Microglia play a key role in the immune response and inflammatory reaction that occurs in response to ischemic stroke. Activated microglia promote neuronal damage or protection in injured brain tissue. Extracellular signals polarize the microglia towards the M1/M2 phenotype. The M1/M2 phenotype microglia released pro- and anti-inflammatory cytokines which induce the activation of neural stem/progenitor cells (NSPCs). In this study, we investigated how the cytokines released by microglia affect the activation of NSPCs. First, we treated BV2 cells with a lipopolysaccharide (LPS; 20 ng/ml) for M1 phenotype microglia and interleukin-4 (IL-4; 20 ng/ml) for M2 phenotype microglia in BV2 cells. Mice were subjected to transient middle cerebral artery occlusion (tMCAO) for 1 h. In ex vivo, brain sections containing the subventricular zone (SVZ) were cultured in conditioned media of M1 and M2 phenotype-conditioned media for 3 d. We measured the expression of cytokines in the conditioned media by RT-PCR and ELISA. The M2 phenotype microglia-conditioned media led to the proliferation and neural differentiation of NSPCs in the ipsilateral SVZ after ischemic stroke. The RT-PCR and ELISA results showed that the expression of TGF-α mRNA was significantly higher in the M2 phenotype microglia-conditioned media. These data support that M2 phenotype microglia-derived TGF-α is one of the key factors to enhance proliferation and neural differntiation of NSPCs after ischemic stroke.

Protective effects of deferasirox and N-acetyl-L-cysteine on iron overload-injured bone marrow

Using an iron overload mouse model, we explored the protective effect of deferasirox (DFX) and N-acetyl-L-cysteine (NAC) on injured bone marrow hematopoietic stem/progenitor cells (HSPC) induced by iron overload. Mice were intraperitoneally injected with 25 mg iron dextran every 3 days for 4 weeks to establish an iron overload (Fe) model. DFX or NAC were co-administered with iron dextran in two groups of mice (Fe+DFX and Fe+NAC), and the function of HSPCs was then examined. Iron overload markedly decreased the number of murine HSPCs in bone marrow. Subsequent colony-forming cell assays showed that iron overload also decreased the colony forming capacity of HSPCs, the effect of which could be reversed by DFX and NAC. The bone marrow hematopoiesis damage caused by iron overload could be alleviated by DFX and NAC.

Relationship between endometrial stem/progenitor cells and gynecological diseases / 中国综合临床

The existence of stern/progenitor cells in the endometrium was postulated many years ago,but the first functional evidence was published in 2004.The study of endometrial stem/progenitor cells had opened up an active field in the subsequent 10 years.Human endometrium has immense regenerative capacity and bilayer structure,in which the upper functionalis is shed at menses and regenerates from the remaining basalis in the subsequent cycle,these characteristics had been the motivation for investigators to identify and characterize endometrial stem/progenitor cell populations.The discovery of specific markers for endometrial stem/progenitor cells has enabled the examination of their role in endometrial proliferative disorders,including endometriosis,adenomyosis and Asherman's syndrome.

Effects of interleukin-6 in promoting corneal epithelial stem/progenitor cell regeneration and accelerating corneal epithelial wound healing in diabetic mouse / 中华实验眼科杂志

Background Interleukin-6 (IL-6) is a pleiotropic cytokine involving in inflammation and wound healing.Previous report found that IL-6 increases phosphorylated STAT3 (p-STAT3) level and promotes corneal epithelial wound healing by stimulating migration.However,the essential role of IL-6 in corneal epithelial wound healing and the expression changes in diabetic mellitus remains unknown.Objective This study was to explore the roles of IL-6 in corneal epithelial proliferation and wound healing in both normal and diabetic mice.Methods Fifty-two normal C57BL/6 mice were randomized into normal control group (32 mice) and diabetic group (20 mice).Type 1 diabetic mellitus was induced by intraperitoneal injections of 50 mg/kg streptozotocin once per day for consecutive 5 days in the mice of the diabetic group.Whole corneal epithelium was scraped in all mice,and the corneal epithelial defect area was examined by fluorescein staining in 24,48 and 72 hours after corneal epithelium removal.Recombinant mouse IL-6 or anti-IL-6 blocking antibody of 5 μl were subconjunctivally injected according to the grouping and contrasted with PBS injection group or isotype control antibody group,respectively.TKE2 cells,a mouse corneal epithelial stem/progenitor cell line,were trypsinized and incubated in the KSFM with different concentrations of IL-6 or without IL-6,and colony formation efficency (CFE) was examined by crystal violet staining.The expressions of △NP63 and Ki67,specific makers of stem cells,were detected by immunofluorescine technology.The expressions of △NP63,Ki67 and p-STAT3 proteins were assayed in the cells by Western blot,respectively.The expression of IL-6 mRNA and protein in the regenerated corneal epithelium was detected by real time quantitative PCR and ELISA.The use and care of the mice complied with the Statement of Association for Research in Vision and Ophthalmology.Results The percentage of residual corneal epithelium defect area with initial detect area was gradually shrinked over time after PBS and IL-6 injection in both normal control mice and diabetic mice,and the percentage of residual corneal epithelium defect area was significantly reduced in the IL-6 injected group compared with the PBS injected group (normal control group:Fgroup =19.982,P ＜ 0.01;Ftime =589.350,P ＜ 0.01;Diabetic group:Fgroup =25.411,P＜0.01;Ftime =334.807,P＜0.01).The CFE was (13.23± 1.12)％,(15.87± 1.30)％,(21.69±1.62)％,(25.33±1.28)％ and (18.67±1.54)％ in the blank control group and 10,20,50,100 ng/ml IL-6-treated groups,respectively,showing a gradual increase of CFE dependent upon IL-6 concetrations (F =35.547,P＜0.01).The expressions of △NP63,Ki67,p-STAT3 proteins in the cells were gradually increased over time after 50 ng/ml IL-6 treated for 5,10,15,30 and 60 minutes,and the relative expression level of the cytokines was significnatly higher in the IL-6 cultured groups than that without IL-6 culture group (all at P＜0.05).The relative expression of IL-6 mRNA in the regenerated corneal epithelilum was 0.45±0.21 and 1.00±0.16 in the diabetic group and normal control group,respectively,and compared with the normal control group,the expression of IL-6 mRNA reduced by 56％ (t=3.42,P=0.03).The content of IL-6 protein in regenerated corneal epithelium of the diabetic group was (257±12) ng/μl,which was significantly lower than (323 ± 17) ng/μl of the normal control group (t =5.60,P＜0.01).Conclusions IL-6 promotes the proliferation and regeneration of corneal limbal stem cells to repair defected corneal epithelium by activating STAT3 signaling pathway in both normal and diabetic mice,while the blocking of endogenous IL-6 impairs the corneal epithelial cell activation and wound healing.

AFT024-SCF construction and functional identification / 中国病理生理杂志

AIM:To construct AFT024-SCF cell line and HPC-Lhx2 cell line for confirming the biological function of AFT024-SCF.METHODS:The HPC-Lhx2 cell line, AFT024-SCF cell line and AFT024-GFP cell line were constructed by retro-viral infection.The expression of stem cell factor(SCF) in AFT024-SCF cells was detected by real-time PCR and Western blot.SCF in the supernatant of AFT024-SCF was detected with ELISA.The supernatant of AFT024-SCF and AFT024-GFP were collected and then diluted (1:10) with basic IMDM medium.So we made 4 culture medium:AFT024-SCF medium was used for experiment group, AFT024-GFP medium was used for endogenous negative control, IM-DM basic medium was used for exogenous negative control, and IMDM basic medium with SCF was used for positive con-trol.SCF-dependent HPC-Lhx2 cell line was cultured in these 4 different medium for 72 h.According to MTT method and colony forming experiment, the biological function of AFT024-SCF was confirmed by the proliferation ability of SCF-depend-ent HPC-Lhx2 cell line.RESULTS:SCF was highly expressed in AFT024-SCF cells.After cultured for 72 h, neither IM-DM basic medium nor GFP-AFT024 medium support HPC-Lhx2 cell line proliferation.However, AFT024-SCF medium supported HPC-Lhx2 cell line expansion as well as the positive control medium.CONCLUSION:AFT024-SCF cells ex-press SCF successfully and recombinant SCF can be replaced by the supernatant of AFT024-SCF culture medium for expan-ding HPC-Lhx2 cell line in vitro.

Protective effects of tannins in sanguisorbae radix on myelosuppression mice / 中草药·英文版

Objective: To explore the protective effects of tannins in Sanguisorba Radix (TSR) on myelosuppression mice induced by cyclophosphamide (CTX). Methods: TSR was ig given at the dose of 20 mg/kg for 10 d after ip administration of CTX (200 mg/kg). Results: TSR could significantly increase the numbers of white blood cells, red blood cells, and platelets of myelosuppression in mice. And it could accelerate bone marrow haemopoietic stem/progenitor cells (HSPCs) in myelosuppression mice and enhance cell proliferation by promoting cell cycles from G0/G1 phase to access into S and G2/M phases, then the reduced number of HSPCs induced by CTX was reversed. Moreover, TSR could increase the mRNA and protein expression levels of O(6)-methylguanine-DNA methyltransferase (MGMT) in HSPCs of myelosuppression mice. Conclsion: TSR has a protective function against CTX-induced myelosuppression. The mechanism might be related to protecting hematopoietic stem cells of bone marrow, stimulating hematopoiesis recovery, as well as preventing the apoptosis of hematopoietic stem cells induced by CTX. © 2014 Tianjin Press of Chinese Herbal Medicines.

Advances of Notch signaling regulating fetal liver stem/progenitor cells: implications for the therapy of end-stage liver diseases / 中华肝胆外科杂志

Cell therapy has a very promising potential for end-stage liver diseases (ESLD).Fetal liver stem/progenitor cells (FLSPCs) have advantages of safety,high survival and proliferation rates,and a small volume,all which make them ideal for liver disease stem cell therapy.During the early phase of our study,we applied a three-step separation method to enrich FLSPCs and obtained a separation efficiency similar to that of the flow-cell sorting method.Additionally,using a fulminant hepatic failure model in rats,we have demonstrated that FLSPCs can contribute to morphological and functional recovery of the liver.This manuscript will discuss how FLSPCs can be induced to accurately differentiate into hepatocytes and cholangiocytes and how FLSPCs maintain self-renewal.The Notch signaling plays a critical role in regulating the differentiation and self-renewal of many types of stem cells.Our previous findings have shown that the Notch signaling plays an important role in FLSPCs differentiation into hepatocytes.Therefore,the Notch signaling might be involved in the differentiation and self-renewal of FLSPCs.We conducted a study on the regulatory effects and relative molecular mechanisms of the Notch signaling on FLSPCs and found the corresponding interfering target,which might become an index for the clinical application of FLSPCs.

Bone marrow stem/progenitor cell mobilization in C57BL/6J and BALB/c mice / 한국실험동물학회지

Bone marrow (BM) has been considered as a reservoir of stem/progenitor cells which are able to differentiate into ectodermal, endodermal, and mesodermal origins in vitro as well as in vivo. Following adequate stimulation, such as granulocyte stimulating factor (G-CSF) or AMD3100, BM resident stem/progenitor cells (BMSPCs) can be mobilized to peripheral blood. Several host-related factors are known to participate in this mobilization process. In fact, a significant number of donors are resistant to G-CSF induced mobilization protocols. AMD3100 is currently used in combination with G-CSF. However, information regarding host-related factors which may influence the AMD3100 directed mobilization is extremely limited. In this study, we were to get some more knowledge on the host-related factors that affect the efficiency of AMD3100 induced mobilization by employing in vivo mobilization experiments. As a result, we found that C57BL/6J mice are more sensitive to AMD3100 but less sensitive to G-CSF which promotes the proliferation of BMSPCs. We excluded S1P as one of the host related factor which influences AMD3100 directed mobilization because pre-treatment of S1P receptor antagonist FTY720 did not inhibit BMSPC mobilization. Further in vitro experiments revealed that BALB/c mice, compared to C57BL/6J mice, have less BMSPCs which migrate in response to host related factors such as sphingosine-1-phosphate (S1P) and to CXCL12. We conclude that AMD3100-directed mobilization depends on the number of BMSPCs rather than on the host-related factors. These results suggest that the combination of AMD3100 and G-CSF is co-operative and is optimal for the mobilization of BMSPCs.

Mechanism of ginsenoside Rg1 against hematopoietic stem/progenitor cells senescence during serial transplantation / 中草药

Objective: To investigate the mechanism of ginsenoside Rg1 against the hematopoietic stem/progenitor cell (HSC/HPC) senescence during serial transplantation. Methods: Male mice Sca-1+HSC/HPC isolated and purified by magnetic activated cell sorting (MACS) were transplanted for three generations to establish aging in vivo model of mice. The female recipient mice radiated lethal dose from 60Co γ ray were divided into four groups: the control, model, Rg1-treated (20 mg/kg), and Rg1 prevention (20 mg/kg) groups. The mice were administered once daily for 30 d. The expression of p16INK4a, p19Arf, p53, and p21Cip1/Waf1 mRNA was detected by quantitative PCR. The expression of p16INK4a, p21Cip1/Waf1, CDK4, CDK2, CyclinD1, and CyclinE proteins was examined by Western blotting. Results: Compared with the model group, the expression of p16INK4a, p19Arf, p53, p21Cip1/Waf1mRNA and p16INK4a, p21Cip1/Waf1, CyclinD1 proteins was down-regulated, and the expression of CDK4, CDK2, and CyclinE proteins was up-regulated in both Rg1-treated and Rg1 prevention groups. The all index changes of mice in Rg1 prevention group were significantly higher than those of mice in Rg1-treated group. Conclusion: Rg1 could prevent and treat the Sca-1+HSC/HPC senescence during serial transplantation in which the signaling pathways of p16INK4a-Rb and p19Arf-p53-p21Cip/Waf1 may play an important role.

Induction of Angiogenesis and Neurogenesis by Serum from Rats Treated with Shunaoxin Dropping Pills / 中草药·英文版

Objective Shunaoxin Dropping Pills (SDPs), a Chinese patent medicine, has been used widely in China for the treatment of headache, amnesia, and insomnia. The aim of the present study is to observe the effect of SDPs on inducing angiogenesis and neurogenesis in vitro. Methods The present testing system using the serum obtained from animals ig treated with SDPs and a co-culture system in vitro was used to investigate if SDPs promotes brain microvascular endothelial cells (BMECs) tube formation and neural differentiation of neural stem/progenitor cells (NSPCs), which plays important roles in angiogenesis and neurogenesis. Results The SDPs serum sampled from rats ig treated with SDPs for 3 d dose-dependently promoted the tube like structure formation of cultured BMECs, and enhanced the fraction of MAP-2 positive cells of NSPCs, which co-cultured with the BMECs and astrocyte. In addition, there was no significant change in the percentage of glial fibrillary acidic protein positive cells. Conclusion Our results show that SDPs serum can induce neural differentiation and BMECs tube formation in vitro.