Human neural stem cells.

'Human neural stem cells' jointly drafted and agreed upon by experts from the Chinese Society for Stem Cell Research, is the first guideline for human neural stem cells (hNSCs) in China. This standard specifies the technical requirements, test methods, test regulations, instructions for use, labelling requirements, packaging requirements, storage requirements, transportation requirements and waste disposal requirements for hNSCs, which is applicable to the quality control for hNSCs. It was originally released by the China Society for Cell Biology on 30 August 2022. We hope that publication of the guideline will facilitate institutional establishment, acceptance and execution of proper protocols, and accelerate the international standardization of hNSCs for clinical development and therapeutic applications.

Development of a CRISPR/Cas9D10A Nickase (nCas9)-Mediated Genome Editing Tool in Streptomyces.

Streptomycetes have a strong ability to produce a vast array of bioactive natural products (NPs) widely used in agriculture and veterinary/human medicine. The recently developed CRISPR/Cas9-based genome editing tools have greatly facilitated strain improvement for target NP overproduction as well as novel NP discovery in Streptomyces. However, CRISPR/Cas9 shows high toxicity to the host, limiting its application in many Streptomyces strains with a low DNA transformation efficiency. In this study, we developed a low-toxicity CRISPR/Cas9D10A nickase (nCas9)-based genome editing tool in the model strain Streptomyces coelicolor M145. We showed that in the presence of both targeting sgRNA and Cas proteins, utilization of nCas9 instead of Cas9 significantly reduced the toxicity to the host and greatly enhanced cell survival. Using this tool, we achieved deletion of single genes and gene clusters with efficiencies of 87-100 and 63-87%, and simultaneous deletion of two genes or gene clusters with efficiencies of 47 and 43%, respectively. The editing efficiency of nCas9 is comparable to that of the Cas9-mediated editing tool. Finally, the nCas9-based editing tool was successfully applied for genome editing in the industrial rapamycin-producing strain Streptomyces rapamycinicus, in which CRISPR/Cas9 cannot work well. We achieved the deletion of three tested genes with an efficiency of 27.2-30%. Collectively, the CRISPR/nCas9-based editing tool offers a convenient and efficient genetic modification system for the engineering of streptomycetes, particularly those with low DNA transformation efficiency.

Calcium isotopic geochemistry of geothermal systems in the tectonically active southeastern Tibetan Plateau.

The global Calcium (Ca) cycle is closely coupled to the carbon cycle, and Ca isotopes have potential in tracing it. Even though groundwater is one of the main reservoirs of Ca at the Earth's surface, few data are available for groundwater, and the behavior of Ca and its isotopes in geothermal systems remains unknown. Here we analysed the stable Ca and radiogenic Sr isotope compositions of thermal waters distributed along the Jinsha and Yalong river valleys in the southeastern Tibetan Plateau. The Ca isotopic composition of the thermal water ranges from 0.45 to 2.16 ‰ (δ44/40Ca values relative to SRM 915a). The thermal waters collected from carbonate aquifers have higher δ44/40Ca values than bedrocks, which was attributed to secondary carbonate precipitation accompanied by CO2 degassing. In contrast, δ44/40Ca values in thermal waters collected from clastic and igneous rocks are similar to bedrock. Despite some thermal waters undergoing secondary silicates formation and CaNa ion exchange, such processes maybe not play a significant role in governing the Ca isotopic composition of these thermal waters. This suggests that Ca isotopes can be used to trace secondary carbonate precipitation driven by CO2 degassing (e.g. travertine) in geothermal systems located in tectonically active areas.

Role of Immune and Inflammatory Mechanisms in Stroke: A Review of Current Advances.

Stroke accounts for a large proportion of morbidity and mortality burden in China. Moreover, there is a high prevalence of the leading risk factors for stroke, including hypertension and smoking. Understanding the underlying mechanisms and developing effective therapeutic interventions for patients with stroke is a key imperative. The pathophysiology of stroke involves a complex interplay between the immune and inflammatory mechanisms. Focal brain inflammation triggered by neuronal cell death and the release of factors such as damage-associated molecular patterns can further exacerbate neuronal injury; in addition, impairment of the blood-brain barrier, oxidative stress, microvascular dysfunction, and brain edema cause secondary brain injury. Immune cells, including microglia and other infiltrating inflammatory cells, play a key role in triggering focal and global brain inflammation. Anti-inflammatory therapies targeting the aforementioned mechanisms can alleviate primary and secondary brain injury in the aftermath of a stroke. Further experimental and clinical studies are required to explore the beneficial effects of anti-inflammatory drugs in stroke.

Transfer characteristic of fluorine from atmospheric dry deposition, fertilizers, pesticides, and phosphogypsum into soil.

Fluorine (F) is widely dispersed in the environment and frequently used in industry and agriculture with a high migration ability. Thus, it is essential to understand the leaching characteristic of F in soil from industry and agriculture sources. Several sources of F pollutants in soil, including fertilizers, pesticides, phosphogypsum, and atmospheric deposition, were selected to investigate leaching characteristics of F in soil by leaching experiments. The addition of phosphate fertilizer and compound fertilizer (N:P:K = 20:10:15) enhanced the leachability of F in soil and the proportion of F leached out from soil treated by these fertilizers were 0.25% and 0.24%, respectively. However, unanticipated lower leachability of F appeared in compound fertilizer (N:P:K = 17:17:17), nitrogen fertilizer, dipterex, fluoroglycofen, fluopimomide, simulative dry deposition (YF3), and phosphogypsum loaded soils compared with additive-absent treatment. Although phosphogysum had a high F concentration, minimum proportion of F released (0.18%) was observed in phosphogypsum-coverd soil. The amounts of F leaching-out from surface soils (0-25 cm) treated with nitrogen fertilizer decreased 1.03 kg ha-1 comparing with blank control. Soil with phosphate fertilizer leached 5.47 kg F ha-1 a year, having the highest environment risk to deeper soil and groundwater. However, phosphogypsum and dry deposition of airbone F chemical had few effects on F leaching in soil. F-containing materials from agricultural process may leach more F from surface soils than industrial sources.

Adsorption-desorption and co-migration of vanadium on colloidal kaolinite.

Vanadium (V) pollution in soil has been widely noted, while knowledge about the effect of soil colloid on migration of V is scarce. Batch adsorption-desorption and transportation of the colloid-adsorbed V in columns packed with quartz sand under various environment conditions were carried out to explore the retention and transportation of V by colloidal kaolinite. Batch adsorption-desorption studies show that the adsorption of V by the colloidal kaolinite was mainly specific adsorption and redox played a limited role in the adsorption process. The maximum adsorption capacity of the colloidal kaolinite was 712.4 mg g-1, and about 5.9-8.7% of the adsorbed V could be desorbed. Both the adsorption-desorption and migration of V with colloidal kaolinite were highly ambient condition dependent. The column studies show that V was highly mobile in the saturated porous media. An easier transfer of V with an increase in pH, IS, and velocity of flow was noted. However, the increase of IS lead to the blockage of the colloidal kaolinite transportation. The recovery rate of the colloidal kaolinite at pH 7 and 9 was 2.0 and 2.1 times that at pH 5, respectively. The migration of colloidal-adsorbed V in sand column preceded that of V ion, but more colloidal-bound V than V ion remained in the column. Lack of consideration of the combination and co-transportation of V and colloidal kaolinite will lead to an overestimation of the risk of V to deeper soil profiles and groundwater. Graphical abstract.

[Research and application progresses on rice water quality (RICEWQ)model.] / ç¨»ç”°æ°´è´¨æ¨¡åž‹ç ”ç©¶å’Œåº”ç”¨è¿›å±•.

Rice production presents a unique environmental condition, with the diseases, insects and weeds being serious during production process. Many kinds of pesticides are used with high frequency. Some pesticides will leach into surrounding water, which shows a high risk for pollution. With the increasing costs of pesticide monitoring and field experiments, mathematical models have become an indispensable part during the process of pesticide registration. As the most reliable and widely used model for pesticide exposure assessment in European paddy fields, rice water quality (RICEWQ) model was mainly used to predict pesticide concentrations in flooded paddy fields and water, and had been preliminarily applied in China. In this review, system structure, the main processes of pesticide fate involved, input and output of RICEWQ model were briefly introduced, and the research progress at home and abroad were summarized. This review would promote the application of RICEWQ model in China and provide reference for related research.

lncRNA HCG11 regulates cell progression by targeting miR-543 and regulating AKT/mTOR pathway in prostate cancer.

Prostate cancer (PCa) is a common cancer worldwide, which mostly occurs in males over the age of 50. Accumulating evidence have determined that long non-coding RNA/microRNA (lncRNA/miRNA) axis plays a critical role in cell progression of cancers, including PCa. However, the pathogenesis of PCa has not been fully indicated. In this study, quantitative real-time polymerase chain reaction was used to detect the expression of HCG11 and miR-543. Western blot was applied to measure the protein expression of proliferating cell nuclear antigen, cleavage-caspase 3 (cle-caspase 3), N-cadherin, E-cadherin, GAPDH, P-AKT, AKT, p-mTOR, and mTOR. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), transwell invasion, and transwell migration assay were used to detect cell proliferation, invasion, and migration, respectively. The function and mechanism of lncRNA HCG11 were confirmed in PCa cell and xenograft mice models. Luciferase assay indicated that miR-543 was a target miRNA of HCG11. Further investigation revealed that overexpression of HCG11 inhibited cell proliferation, invasion, and migration, whereas induced cell apoptosis by regulating miR-543 expression in vitro and in vivo. More than that, lncRNA HCG11 inhibited phosphoinositide-3 kinase/protein kinaseB (PI3K/AKT) signaling pathway to suppress PCa progression. Our data showed the overexpression of HGC11-inhibited PI3K/AKT signaling pathway by downregulating miR-543 expression, resulting in the suppression of cell growth in PCa. This finding proved a new regulatory network in PCa and provided a novel therapeutic target of PCa.

Distribution, health risk assessment, and anthropogenic sources of fluoride in farmland soils in phosphate industrial area, southwest China.

The high concentration of fluoride (F) in soils has become a rising concern for its toxicity to microbes, plants, animals and human health. In the present study, the spatial and vertical distribution, health risk assessment and anthropogenic sources of F in farmland soils in an industrial area dominated by phosphate chemical plants were studied. Concentrations of total fluoride (TF) and water soluble fluoride (WSF) in the surface soils decreased with distance within the range of 2500 m at the prevailing downwind of the industrial area. The soil TF and WSF concentrations in 0-40 cm profiles were higher than those in 40-100 cm layers in the industrial area. At the prevailing downwind of the industrial area within 700 m, the hazard quotient values of human exposure to surface soils were higher than 1, indicating that a potential risk may exist for human health in this area. The main exposure pathway for children and adults was oral ingestion and particulate inhalation, respectively. The source apportionment model of soil F was modified based on years' historical data and experimental data. The results showed that the proportion of anthropogenic sources of soil F was dustfalls (69%) > irrigation water (23%) > air (5%) > chemical fertilizers (3%) in the industrial area. The high F concentration of dustfalls was mainly due to the phosphate rock, phosphogypsum, and surface soils with high F contents from the factories. In order to safeguard human health and alleviate hazards of F to surroundings, the control of pollutants emission from factories was a basic and vital step to reduce F in the soils in industrial areas.

Vertical distribution of fluorine in farmland soil profiles around phosphorous chemical industry factories.

High concentration of fluorine (F) in agricultural soils has got significant attention considering its impacts on human health, but little information was available about F distribution in farmland soil profiles around phosphorous chemical industry factories. In present study, farmland soil profiles and relevant medium samples were collected from farmlands around a main phosphorous chemical base in southwest China. At 0-100-cm profiles, concentrations of soil total F (Ft, 400.9-1612.0 mg kg-1) and water soluble F (Fw, 3.4-26.0 mg kg-1) decreased with profile depth in industrial areas. Industrial activities enhanced F concentration in soil mainly at 0-40-cm profiles. No disparity for both Ft and Fw distributions in paddy-dry land rotation field and dry land indicates short-term land utilization could not affect the F distribution in soil profiles. Correlation analysis showed soil organic matter and wind direction were important factors influencing the distribution of F in soil profiles. The shutdown of factory and government control of industrial emissions effectively decreased the ambient air F (Fa) concentrations in industrial areas. In where Fa and dustfall F concentrations were high, high soil Ft, Fw, and crop edible part F concentrations were found.

Effect of lead on plant availability of phosphorus and potassium in a vegetable-soil system.

Two typical red soils were sequentially cultivated with celery (Apium graveolens L.) and Chinese cabbage (Brassica chinensis L.) in a greenhouse to determine the effect of lead (Pb) on plant availability of phosphorus (P) and potassium (K) in the soils. The concentrations of available P as estimated by the 0.05 mol L-1 HCl-0.025 mol L-1 (1/2 H2SO4) extraction and available K estimated by the NH4OAc extraction method in the crop-free soils were not affected by Pb treatment. Plant P concentrations in the above-ground part of celery and Chinese cabbage exposed to Pb were either lower or showed no significant difference to the control.

Anticancer properties of Semen Euphorbiae towards ACHN human renal adenocarcinoma cells by inducing apoptosis.

The main objective of the present research work was to evaluate the antitumor effects of ethanol extract of Semen Euphorbiae (EESE) in ACHN human renal carcinoma cells. The effects on apoptosis induction, cell cycle phase distribution and livin protein expression were also evaluated. MTT assay was used to assess cytotoxic effects of the extract while as clonogenic assay was used to evaluate effects on colony formation tendency. Inverted phase contrast and fluorescence microscopic techniques were used to evaluate effects of EESE on cellular morphology and apoptosis. Flow cytometry using annexin V-FITC and propidium iodide were used to quantify the extent of apoptosis and also to evaluate effects on cell cycle. Results indicate that ethanol extract of Semen Euphorbiae exhibited potent cytotoxic effects in ACHN human renal cancer cells. These effects were found to be dose-dependent as well as time dependent. Clonogenic assay revealed that EESE led to dose-dependent inhibition of colony formation in these cells. EESE-treated cells also showed evident signs of alterations and deformations in cell morphology including detachment of cells from one another forming small cluster of cells. In contrast to untreated control cells, EESE-treated cells with 10, 100 and 200 µg/ml dose showed an increase in the number of cells emitting red/orange fluorescence indicating onset and execution of apoptosis. EESE extract also led to G2/M cell cycle arrest in these cells.

Comparative evaluation of copeptin and NT-proBNP in patients with severe acute decompensated heart failure, and prediction of adverse events in a 90-day follow-up period: A prospective clinical observation trial.

The present study compared the prognostic value of a marker, the C-terminal section of the arginine vasopressin prohormone (copeptin), with N-terminal B-type natriuretic peptide (NT-proBNP) in patients with severe acute decompensated heart failure. A prospective, observational cohort study was conducted in a tertiary care hospital and enrolled 129 patients with severe acute decompensated heart failure. Clinicians were blinded to investigational markers except NT-proBNP, and the study participants were followed up for 90 days. The end-point was a composite of cardiovascular death or re-hospitalization due to decompensated heart failure. Of the 129 patients enrolled, 47 reached the end-point and 82 were in a stable condition during follow-up. Receiver operating characteristic curve analysis revealed that the areas under curve for the prediction of adverse events within 90 days were similar for copeptin [0.602±0.052; 95% confidence interval (CI), 0.499-0.705], NT-proBNP (0.659±0.048; 95% CI, 0.565-0.753) and their combination (0.670±0.050; 95% CI, 0.573-0.767). Kaplan-Meier survival analysis showed that the predictive value of NT-proBNP regarding the probability of survival was superior compared with that of copeptin (log-rank test for trend, P=0.001 vs. 0.040). Furthermore, multivariate Cox proportional-hazards regression analysis revealed that increased NT-proBNP and copeptin plasma concentrations were significant independent predictors of adverse events. The present study provided evidence that copeptin has similar predictive properties compared with NT-proBNP regarding adverse events within 90-days in patients with severe acute decompensated heart failure, but that copeptin may not provide superior 90-day prediction compared to NT-proBNP.

[Differentiation potential of CD41⁺ cells derived from the mouse aorta-gonad-mesonephros region, yolk sac and embryonic circulating blood].

OBJECTIVE: To compare the differentiation ability difference of hematopoietic, mesenchymal and endothelial potential between CD41⁺ cells derived from the mouse aorta-gonadmesonephros (AGM) region, yolk sac (YS) and embryonic circulating blood (CB). METHODS: CD41⁺ cells were sorted from AGM, YS and CB. The CD45 and c-kit expression were studied in CD41⁺ cells by flow cytometry. IL-3 and bone morphogenetic protein 4 (BMP-4) treatment together with semi solid culture were used to assess hematopoietic potential difference of CD41⁺ cells. Immunofluorescence staining of α-SMA was used to assess mesenchymal potential difference. The endothelial cell induction system was used to assess endothelial potential difference. RESULTS: The proportions of CD45+ cells in CD41⁺ population were 51.9% (AGM), 45.8% (YS) and 22.2% (CB), respectively, while those of c-kit⁺ cells were 40.0% (AGM), 39.6% (YS) and 36.2% (CB), respectively. After stimulated by IL-3 factor, the number of total colonies increased in all three groups-derived CD41⁺ cells compared to that of unstimulated group[(14.1±1.9) vs (1.2±0.2), (32.4±1.1) vs (18.4±2.2) and (41.8±0.9) vs (10.4±1.8)], (P<0.01). After stimulated by BMP-4 factor, compared to unstimulated group, CFU-Mix colony number in CD41⁺ cells from AGM region and YS were significantly decreased[(0.5±0.6) vs (3.2±0.8), (1.3±0.7) vs (7.4±1.7)](P<0.01), but there was no difference in CB group[(2.5±0.5) vs (3.9±1.5)](P>0.01). The mesenchymal marker α-SMA was highly expressed in CD41⁺ cells from AGM region and YS, but lowly expressed in CD41⁺ cells from CB. CONCLUSION: There are some differences between CD41⁺ cells in AGM region, YS and CB on hematopoietic cell surface marker expression, hematopoietic colony formation with IL-3 and BMP-4 stimulation.

[Analyzing hematopoietic development of mouse embryonic AGM region by tissue culture].

To analyze hematopoietic kinetics of mouse embryonic aorta-gonad-mesonephros (AGM) region, an in vitro tissue culture method was developed in this study, partly based on previous reports. After 2 days of tissue culture, a significant number of erythro myeloid progenitors, as quantitated by colony forming assay was detected in the AGM region. Moreover, the cells from cultured E10.5 AGM region could generate 10.8 ± 3.5 colony-forming unit in spleen (CFU-S) per tissue on average. Transplantation of cultured E10.5-E11.0 AGM cells resulted in efficient (85.7% repopulated) and long-term (> 4 months) reconstitution of lethally irradiated adult recipients with remarkable chimerism [(51.12 ± 21.17)%]. The multilineage contribution of donor cells was validated by significant engraftment of myeloid and/or lymphoid cells in peripheral blood, bone marrow, spleen and thymus of recipients. Taken together, the tissue culture method can enable us to manipulate the AGM region in vitro, fulfilling a systematic evaluation of developmental kinetics of various hematopoietic precursor cells, particularly HSC, in normal and mutant mid-gestation mouse embryos.

Migration of dorsal aorta mesenchymal stem cells induced by mouse embryonic circulation.

Mesenchymal stem cells (MSCs) represent powerful tools for regenerative medicine for their differentiation and migration capacity. However, ontogeny and migration of MSCs in mammalian mid-gestation conceptus is poorly understood. We identified canonical MSCs in the mouse embryonic day (E) 11.5 dorsal aorta (DA). They possessed homogenous immunophenotype (CD45(-)CD31(-)Flk-1(-)CD44(+)CD29(+)), expressed perivascular markers (α-SMA(+)NG2(+)PDGFRß(+)PDGFRα(+)), and had tri-lineage differentiation potential (osteoblasts, adipocytes, and chondrocytes). Of interest, MSCs were also detected in E12.5-E13.5 embryonic circulation, 24 hr later than in DA, suggesting migration like hematopoietic stem cells. Functionally, E12.5 embryonic blood could trigger efficient migration of DA-MSCs through platelet-derived growth factor (PDGF) receptor-, transforming growth factor-beta receptor-, but not basic fibroblast growth factor receptor-mediated signaling. Moreover, downstream JNK and AKT signaling pathway played important roles in embryonic blood- or PDGF-mediated migration of DA-derived MSCs. Taken together, these results revealed that clonal MSCs developed in the mouse DA. More importantly, the embryonic circulation, in addition to its conventional transporting roles, could modulate migration of MSC during early embryogenesis.

[Clarification of lymphoid potential in mouse E8.5 embryos by OP9/OP9-DL1 coculture].

The anatomical location of lymphocyte ontogeny in the developing mouse embryo remains controversial. To define the site that can generate lymphocytes de novo, the intraembryonic splanchnopleura (SP) and extraembryonic yolk sac (YS) at 8.5 days postcoitum, when systemic circulation is not established, were investigated. The results indicated that in standard colony forming assay, the cells from both splanchnopleura and yolk sac formed typical myeloerythroid colonies, but their types were distinct. When cocultured with the OP9, the splanchnopleura produced B cells expressing B220, CD19 and surface IgM. Using a three-step culture protocols with the OP9 expressing Delta-like 1 as feeders, the splanchnopleura produced immature T precursor cells (CD44-/CD25+) and more mature single positive T cells (CD4+/CD8-) after 16 days of incubation. However, the yolk sac failed to generate B and T lymphocytes under identical conditions. It is concluded that prior to linked embryonic circulation, the splanchnopleura other than the yolk sac had robust lymphoid potential in vitro. In the future, more reliable evidence from novel model animals will ultimately delineate the embryonic origin of lymphocytes in vivo.

Interleukin-3 promotes hemangioblast development in mouse aorta-gonad-mesonephros region.

BACKGROUND: The hemangioblast is a bi-potential precursor cell with the capacity to differentiate into hematopoietic and vascular cells. In mouse E7.0-7.5 embryos, the hemangioblast can be identified by a clonal blast colony-forming cell (BL-CFC) assay or single cell OP9 co-culture. However, the ontogeny of the hemangioblast in mid-gestation embryos is poorly defined. DESIGN AND METHODS: The BL-CFC assay and the OP9 system were combined to illustrate the hemangioblast with lymphomyeloid and vascular potential in the mouse aorta-gonad-mesonephros region. The colony-forming assay, reverse transcriptase polymerase chain reaction analysis, immunostaining and flow cytometry were used to identify the hematopoietic potential, and Matrigel- or OP9-based methods were employed to evaluate endothelial progenitor activity. RESULTS: Functionally, the aorta-gonad-mesonephros-derived BL-CFC produced erythroid/myeloid progenitors, CD19(+) B lymphocytes, and CD3(+)TCRbeta(+) T lymphocytes. Meanwhile, the BL-CFC-derived adherent cells generated CD31(+) tube-like structures on OP9 stromal cells, validating the endothelial progenitor potential. The aorta-gonad-mesonephros-derived hemangioblast was greatly enriched in CD31(+), endomucin(+) and CD105(+) subpopulations, which collectively pinpoints the endothelial layer as the main location. Interestingly, the BL-CFC was not detected in yolk sac, placenta, fetal liver or embryonic circulation. Screening of candidate cytokines revealed that interleukin-3 was remarkable in expanding the BL-CFC in a dose-dependent manner through the JAK2/STAT5 and MAPK/ERK pathways. Neutralizing interleukin-3 in the aorta-gonad-mesonephros region resulted in reduced numbers of BL-CFC, indicating the physiological requirement for this cytokine. Both hematopoietic and endothelial differentiation potential were significantly increased in interleukin-3-treated BL-CFC, suggesting a persistent positive influence. Intriguingly, interleukin-3 markedly amplified primitive erythroid and macrophage precursors in E7.5 embryos. Quantitative polymerase chain reaction analysis demonstrated declined Flk-1 and elevated Scl and von Willebrand factor transcription upon interleukin-3 stimulation, indicating accelerated hemangiopoiesis. CONCLUSIONS: The hemangioblast with lymphomyeloid potential is one of the precursors of definitive hematopoiesis in the mouse aorta-gonad-mesonephros region. Interleukin-3 has a regulatory role with regards to both the number and capacity of the dual-potential hemangioblast.

[Current status of study on embryonic hematopoietic development in aorta-gonad-mesonephros -- review].

Aorta-gonad-mesonephros (AGM) is well known as a main structure that de novo generates hematopoietic primary stem cells (HSC) in mid-gestation mammalian embryos. Hemogenic endothelium, and recently, subendothelial mesenchyme as well as hemangioblast are shown as contributing to blood formation in AGM region. AGM-HSC displays dynamic changes in surface markers, including CD41, CD45 and several endothelial-specific molecules. The novel finding of interleukin-3 as a potent regulator of AGM-HSC seems very interesting. Moreover, zebra fish model reveals PGE2 as a novel stimulator of HSC in AGM and kidney marrow, which is also the case in mouse hematopoietic tissues. Identification of mesenchymal stem cells with significant hematopoietic supporting capacity in AGM region suggests an alternative pathway to explore new molecules governing embryonic and adult hematopoiesis. In this paper, the hemogenic model in AGM region, surface markers on HSCs in AGM region and regulation of HSCs in AGM region were reviewed.

[Separation of immortalized mesenchymal stem cell like stromal cells of mouse embryonic aorta-gonad-mesonephros region and their biological characteristics].

To investigate the effects of microenvironment of aorta-gonad-mesonephros (AGM) on embryonic hematopoiesis, mesenchymal stem cell like stromal cells (MSC like stromal cells) derived from dorsal aorta (DA) in AGM region were separated and identified by their growth characteristics, related molecules expression and mesenchymal lineage potentials. Stromal cells from DA region in mouse embryos (E11.5) were isolated and cultured in vitro. After transfected by pSV3neo-SV40, the clones with G418 resistance were selected, and their growth characteristics were studied. The related molecules were analyzed by flow cytometry, and each clone was induced to differentiate into adipocytes, osteocytes, and chondrocytes. The results showed that most clones (20 clones) selected in the mouse DA region held the morphology of fibroblastoid cells. mDAF3 and mDAF18 could be grown in culture for more than 50 passages with G418 resistance, both have the potential to differentiate into adipocytes, osteocytes, and chondrocytes. At the logarithmic growth period, the cell population doubling time is about 24 hours. Surface markers, such as CD29, CD44, CD105 and Sca-1 were positively detected, while low levels of CD34, CD45, and CD31 were detected. It is concluded that immortalized mDAF3 and mDAF18 have the specific phenotype and differential potency of MSC, which suggests that MSC maybe exist in mouse embryonic DA region, where the MSC like stromal cells can be used as a cell model for further research on the modulation activity of DA microenvironment for embryonic hematopoiesis.