Na?ve-like conversion enhances the difference in innate in vitro differentiation capacity between rabbit ES cells and iPS cells

Quality evaluation of pluripotent stem cells using appropriate animal models needs to be improved for human regenerative medicine. Previously, we demonstrated that although the in vitro neural differentiating capacity of rabbit induced pluripotent stem cells (iPSCs) can be mitigated by improving their baseline level of pluripotency, i.e., by converting them into the so-called “naïve-like” state, the effect after such conversion of rabbit embryonic stem cells (ESCs) remains to be elucidated. Here we found that naïve-like conversion enhanced the differences in innate in vitro differentiation capacity between ESCs and iPSCs. Naïve-like rabbit ESCs exhibited several features indicating pluripotency, including the capacity for teratoma formation. They differentiated into mature oligodendrocytes much more effectively (3.3–7.2 times) than naïve-like iPSCs. This suggests an inherent variation in differentiation potential in vitro among PSC lines. When naïve-like ESCs were injected into preimplantation rabbit embryos, although they contributed efficiently to forming the inner cell mass of blastocysts, no chimeric pups were obtained. Thus, in vitro neural differentiation following naïve-like conversion is a promising option for determining the quality of PSCs without the need to demonstrate chimeric contribution. These results provide an opportunity to evaluate which pluripotent stem cells or treatments are best suited for therapeutic use.     

Cell-free extract from porcine induced pluripotent stem cells can affect porcine somatic cell nuclear reprogramming

Pretreatment of somatic cells with undifferentiated cell extracts, such as embryonic stem cells and mammalian oocytes, is an attractive alternative method for reprogramming control. The properties of induced pluripotent stem cells (iPSCs) are similar to those of embryonic stem cells; however, no studies have reported somatic cell nuclear reprogramming using iPSC extracts. Therefore, this study aimed to evaluate the effects of porcine iPSC extracts treatment on porcine ear fibroblasts and early development of porcine cloned embryos produced from porcine ear skin fibroblasts pretreated with the porcine iPSC extracts. The Chariot^TM reagent system was used to deliver the iPSC extracts into cultured porcine ear skin fibroblasts. The iPSC extracts-treated cells (iPSC-treated cells) were cultured for 3 days and used for analyzing histone modification and somatic cell nuclear transfer. Compared to the results for nontreated cells, the trimethylation status of histone H3 lysine residue 9 (H3K9) in the iPSC-treated cells significantly decreased. The expression of Jmjd2b, the H3K9 trimethylation-specific demethylase gene, significantly increased in the iPSC-treated cells; conversely, the expression of the proapoptotic genes, Bax and p53, significantly decreased. When the iPSC-treated cells were transferred into enucleated porcine oocytes, no differences were observed in blastocyst development and total cell number in blastocysts compared with the results for control cells. However, H3K9 trimethylation of pronuclear-stage-cloned embryos significantly decreased in the iPSC-treated cells. Additionally, Bax and p53 gene expression in the blastocysts was significantly lower in iPSC-treated cells than in control cells. To our knowledge, this study is the first to show that an extracts of porcine iPSCs can affect histone modification and gene expression in porcine ear skin fibroblasts and cloned embryos.     

不同杀菌剂对当归主要病害的防效及经济效益比较

针对甘肃当归病害逐年加重态势, 采用田间随机区组试验, 分析比较70%噁霉灵WP?50%氯溴异氰尿酸WP和80%代森锰锌WP对当归主要病害防效及经济效益, 以期筛选出最佳防控药剂?结果表明:3种杀菌剂均能不同程度防治当归病害, 其中以50%氯溴异氰尿酸WP 1.86 g/L对褐斑病?麻口病防效最佳, 分别为70.38%~78.75%?84.48%~91.04%, 70%噁霉灵WP 0.34 g/L和80%代森锰锌WP 7.92 g/L次之, 对褐斑病防效分别为59.14%~63.72%?58.14%~62.84%, 对麻口病防效分别为78.61%~83.57%?78.97%~83.57%?50%氯溴异氰尿酸WP 1.86 g/L能很好地抑制当归抽薹, 抑制率达58.63%~63.15%?3种杀菌剂均能明显提高当归药材产量构成因子, 以80%代森锰锌WP 7.92 g/L处理增产增收效果最好, 两年平均增产量高达5 559.91 kg/hm2, 平均经济效益为194 136.65元/hm2, 但投入产出比最低, 为1∶480.63?50%氯溴异氰尿酸WP 1.86 g/L对当归病害防效及抽薹抑制率最高, 投入产出比为1∶634.02, 在此基础上综合考虑病害防效和经济效益, 50%氯溴异氰尿酸WP 1.86 g/L可作为生产上防控当归主要病害的主推药剂?     

滴灌施肥周期和毛管布设方式对苹果树细根直径时空分布的影响

为探明不同滴灌施肥策略对苹果树细根直径的调控效应,于2019—2021年开展二因素二水平完全组合设计田间试验,毛管布设方式设置一行一管和一行两管,施肥周期设置15 d和30 d,采用微根管原位监测技术,分苹果树正南、正西及东北3个方位和0～19、19～38、38～57、57～76 cm不同深度土层,持续观测苹果树活跃生长期内细根直径的动态变化,分析了苹果树细根直径对毛管布设方式和施肥周期的响应。结果表明：细根直径主要集中在0.5～1.5 mm范围内,约占90%,0～0.5 mm和1.5～2.0 mm直径的细根占比很少。在夏季之前,直径≤1.0 mm的细根增多;夏季之后,细根直径加粗,直径>1.0 mm的细根迅速增加。相较于施肥周期15 d,施肥周期30 d在2020年6—11月和2021年4—7月均能增加细根直径;在大部分土层中,施肥周期30 d会增加细根直径,施肥周期15 d会减小细根直径;在2020年正南和东北方向上,施肥周期15 d会减小细根直径,施肥周期30 d会增加细根直径。一行一管较一行两管在2020年和2021年8月均能增加细根直径;在浅中层土壤(0～38 cm土层...     

甘肃箭筈豌豆种质资源评价

通过对甘肃42份箭筈豌豆(Vicia sativa)种质资源进行研究评价,以期为甘肃绿肥种植筛选优良的箭筈豌豆品种。结果表明,42份箭筈豌豆种质资源各性状指标存在明显差异,其中单株荚数和单株粒数是构成箭筈豌豆产量的主要要素。籽粒产量、鲜草产量、单株粒数和单株荚数4个指标可以反映箭筈豌豆性状的优劣。通过籽粒产量、鲜草产量、单株粒数和单株荚数4项指标进行聚类,可以分为4类,Ⅰ、Ⅱ类相对较好,共有7份材料,籽粒和鲜草产量分别在2 623.26~2 990.25和32 357.70~35 967.38 kg·hm-2,其中苏箭3号、陇箭1号和333/A属于早熟品种,麦类作物收获后的套复种、玉米(Zea mays)前期间作、马铃薯(Solanum tuberosum)绿肥间作、果树绿肥间作和单种绿肥5种利用模式均适宜;山西春箭碗和MB5/794属于中熟品种,则适宜与马铃薯间作、果树间作和单作;西牧820和波兰箭碗属于晚熟品种,则只适宜与果树间作和单作两种利用方式。     

解读涉牧违法行为涉嫌犯罪的主要罪名

畜牧兽医部门执法人员在行政执法过程中对涉牧违法行为是否涉嫌犯罪和涉嫌何种犯罪的把握,是能否依法、及时向刑事司法机关移送案件、加大对涉牧违法犯罪的打击力度的关键因素。本文依据《中华人民共和国刑法》、相关司法解释和相关法律法规,对畜牧兽医部门在行政执法过程中可能遇到的涉嫌犯罪案件3大类12个主要罪名进行了归纳和解读,可为畜牧兽医行政执法更好地与刑事司法衔接提供参考。     

SCINTIGRAPHIC TRACKING OF MESENCHYMAL STEM CELLS AFTER PORTAL,SYSTEMIC INTRAVENOUS AND SPLENIC ADMINISTRATION IN HEALTHY BEAGLE DOGS

Mesenchymal stem cells have been proposed to treat liver disease in the dog. The objective of this study was to compare portal, systemic intravenous and splenic injections for administration of mesenchymal stem cells to target the liver in healthy beagle dogs. Four healthy beagle dogs were included in the study. Each dog received mesenchymal stem cells via all three delivery methods in randomized order, 1 week apart. Ten million fat‐derived allogeneic mesenchymal stem cells labeled with Technetium‐99m (99mTc)‐hexamethyl‐propylene amine oxime(HMPAO) were used for each injection. Right lateral, left lateral, ventral, and dorsal scintigraphic images were obtained with a gamma camera equipped with a low‐energy all‐purpose collimator immediately after injection and 1, 6, and 24 h later. Mesenchymal stem cells distribution was assessed subjectively using all four views. Pulmonary, hepatic, and splenic uptake was quantified from the right lateral view, at each time point. Portal injection resulted in diffuse homogeneous high uptake through the liver, whereas the systemic intravenous injection led to mesenchymal stem cell trapping in the lungs. After splenic injection, mild splenic retention and high homogeneous diffuse hepatic uptake were observed. Systemic injection of mesenchymal stem cells may not be a desirable technique for liver therapy due to pulmonary trapping. Splenic injection represents a good alternative to portal injection. Scintigraphic tracking with 99mTc‐HMPAO is a valuable technique for assessing mesenchymal stem cells distribution and quantification shortly after administration. Data obtained at 24 h should be interpreted cautiously due to suboptimal labeling persistence.     

不同矮化中间砧对金冠苹果树体及根系生长发育的影响

本试验以嫁接不同矮化中间砧的2年生盆栽金冠苹果树为试材,研究不同矮化砧木对金冠树体及根系生长发育的影响。结果表明,不同矮化中间砧影响金冠苹果树体及根系的生长发育。中间砧为GM256的金冠苹果,树体高度及新梢生长量最大,除根系直径外,其余根系形态指标也均为最大值,树体矮化效果最差;SH1树体矮化效果最好;SH6矮化中间砧木与接穗亲和性最好,辽砧2号组合的亲和性最差;d≦0.5mm根径级的毛细根为金冠苹果幼树根系中的主要组成部分。     

The Potential of Mesenchymal Stem Cell in Prion Research

Scrapie and bovine spongiform encephalopathy are fatal neurodegenerative diseases caused by the accumulation of a misfolded protein (PrP^res), the pathological form of the cellular prion protein (PrP^C). For the last decades, prion research has greatly progressed, but many questions need to be solved about prion replication mechanisms, cell toxicity, differences in genetic susceptibility, species barrier or the nature of prion strains. These studies can be developed in murine models of transmissible spongiform encephalopathies, although development of cell models for prion replication and sample titration could reduce economic and timing costs and also serve for basic research and treatment testing. Some murine cell lines can replicate scrapie strains previously adapted in mice and very few show the toxic effects of prion accumulation. Brain cell primary cultures can be more accurate models but are difficult to develop in naturally susceptible species like humans or domestic ruminants. Stem cells can be differentiated into neuron‐like cells and be infected by prions. However, the use of embryo stem cells causes ethical problems in humans. Mesenchymal stem cells (MSCs) can be isolated from many adult tissues, including bone marrow, adipose tissue or even peripheral blood. These cells differentiate into neuronal cells, express PrP^C and can be infected by prions in vitro. In addition, in the last years, these cells are being used to develop therapies for many diseases, including neurodegenerative diseases. We review here the use of cell models in prion research with a special interest in the potential use of MSCs.     

Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells

Background

There is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. Mesenchymal stem cells (MSCs) have been isolated and characterized from various species, but are poorly characterized in goats.

Results

Goat MSCs isolated from bone marrow (BM-MSCs) and adipose tissue (ASCs) have the ability to undergo osteogenic, adipogenic and chondrogenic differentiation. Cytochemical staining and gene expression analysis show that ASCs have a greater capacity for adipogenic differentiation compared to BM-MSCs and fibroblasts. Different methods of inducing adipogenesis also affect the extent and profile of adipogenic differentiation in MSCs. Goat fibroblasts were not capable of osteogenesis, hence distinguishing them from the MSCs. Goat MSCs and fibroblasts express CD90, CD105, CD73 but not CD45, and exhibit cytoplasmic localization of OCT4 protein. Goat MSCs can be stably transfected by Nucleofection, but, as evidenced by colony-forming efficiency (CFE), yield significantly different levels of progenitor cells that are robust enough to proliferate into colonies of integrants following G418 selection. BM-MSCs expanded over increasing passages in vitro maintained karyotypic stability up to 20 passages in culture, exhibited an increase in adipogenic differentiation and CFE, but showed altered morphology and amenability to genetic modification by selection.

Conclusions

Our findings provide characterization information on goat MSCs, and show that there can be significant differences between MSCs isolated from different tissues and from within the same tissue. Fibroblasts do not exhibit trilineage differentiation potential at the same capacity as MSCs, making it a more reliable method for distinguishing MSCs from fibroblasts, compared to cell surface marker expression.

Electronic supplementary material

The online version of this article (doi:10.1186/2049-1891-6-1) contains supplementary material, which is available to authorized users.