Bone marrow-derived stromal cells and cartilage tissue engineering

Articular cartilage repair is one of the most challenging issues which remains to be resolved in clinic work. Discovering of bone marrow stromal cells (BMSC) and its application in tissue engineering provide new methods for the treatment of cartilage defects. High seeding density, appropriate cytokines and three-dimensional culture play important roles in the process of inducing BMSC differentiating into chondrocytes, suitable scaffold is also essential in reconstructing cartilage in vitro by methods of tissue engineering.     

The transdifferentiation of Sca-1+ cells from murine fetal liver

AIM:To explore transdifferentiation potential of Sca-1⁺ cells from murine fetal liver. METHODS:2×10³ of Sca-1⁺ cells from male murine fetal liver were transfused into female mouse irradiated lethally with γ ray from ⁶⁰ Co source (10 Gy) via tail vein. Two months later, FISH and immunohistochemistry were used to detect the situation for transdifferentiating of the donor cells (male cells) in tissues of female recipient mouse. RESULTS:The renal tubular epitheliocyte-like and neurocyte-like cells with Y chromosome were found on the sections of renal and brain tissues from female recipient mice. These cells have phenotype characteristics of RCA⁺/CD₄₅^-F_4/80^- and NueN⁺/CD₄₅^-F_4/80^-, respectively. CONCLUSION:The evidence is provided for Sca-1⁺ cells from murine fetal liver to transdifferentiate into both renal and brain tissue cells.     

Comparison of different conditions inducing embryonic stem cells

AIM: To evaluate the different conditions inducing mouse embryonic stem cells (ESC) in vitro to differentiate into cardiomyocytes. METHODS: BRL conditioned medium was used to promote the growth of ESC and maintain them in an undifferentiated state. During the inducing process, retinoic acid (RA), DMSO, activin-A and TGF-β₁ were used as inducing reagents, and made up six kinds of differentiating medium. Then a three-step method inducing ESC cultured in hanging drops, in suspension and in plating was used to induce the differentiation of ESC. RESULTS: ESC were induced in vitro to differentiate into cardiomyocytes. Of all groups, the highest differentiating rate was observed in the group induced by activin-A (20 μg/L) and TGF-β₁ (2 μg/L). CONCLUSION: The inducing conditions including activin-A (20 μg/L) and TGF-β₁ (2 μg/L) is very valuable in inducing ESC differentiation into cardiomyocytes.     

Bone marrow mesenchymal stem cells are induced to differentiate into cardiomyocytes by hepatocyte growth factor in vitro

AIM: To explore a new method of hepatocyte growth factor (HGF) inducing bone marrow mesenchymal stem cells (MSC) to differentiate into cardiomyocytes. METHODS: Bone marrow MSC was cultured with DMEM media (10% fetal calf serum) 4-6 passages, and induced by HGF (10 μg/L) for 30 d. Automatical beating of the differentiated cells was observed daily with transverse microscopy, or under condition of 0.1% isoproterenol or cal-cium-deprived incubation. Specific cardiac myosin in the cells was indentified by immunochemistry. RESULTS: At 14-20 d of differentiation, bone marrow mesenchymal stem cells formed clones, in 10%-50% of which spontaneous beating cell-mass had come to continuously exist. Isoproterenol increased the beating rate and calcium-deprived media inhibited the beating. The cells were identified to be cardiomyocytes by expression of cardiac myosin heavy chain. CONCLUSION: HGF may induce bone marrow mesenchymal stem cells into cardiomyocytes with high efficiency, but the differentiating pathway of stem cells remains to be further studied.     

提高水稻愈伤组织再生频率的研究

以长时间继代的水稻成熟胚愈伤组织为试验材料,研究激素配比、继代方式、干燥处理和添加物对分化的影响。结果表明,分化培养基中加3 mg/L BA对分化有利;继代培养基中加入3 mg/L ABA,以麦芽糖代替蔗糖,或以甘露醇代替部分蔗糖在一定程度上可以改善愈伤组织状态,提高分化率;干燥处理能明显提高愈伤组织分化率;分化培养中添加一定浓度的AgNO3和CuSO4有利于愈伤组织分化。     

青花菜组培中过氧化物酶及吼哚乙酸氧化酶与形态发生的关系

青花菜在脱分化及分化过程中其过氧化物酶活性升高;脱分化培养的第12天该酶活性的大幅度增加与愈伤组织的明显发生相对应;分化培养的第21天该酶活性的大幅度增加与真正的芽发生相关.脱分化时过氧化物酶同工酶的c带、e带发生在愈伤组织产生以前,可能是脱分化的原团;分化时c带在芽点出现以前的重新出现可能与分化的启动有关.在脱分化过程中吲哚乙酸氧化酶活性略有升高;在分化过程中该酶活性基本保持稳定.     

刺葡萄愈伤组织的诱导和分化培养

以刺葡萄叶片为材料,在1／2MS培养基上附加不同种类及浓度配比的植物激素,研究叶片愈伤组织的诱导及分化培养。实验结果表明：叶片愈伤组织的诱导较容易,最佳诱导培养基配方为以ZT1．0mg／L＋2,4-D2．0mg／L,诱导率可达100％,光培养条件下刺葡萄叶片愈伤组织的诱导率比暗培养的诱导率高。继代培养愈伤组织能诱导出根,但诱导出芽不易,刺葡萄叶片再生植株较难。增殖培养ZT以1．0mg／L＋NAA0．01mg／L为佳,1周增殖率可达1．292％。     

温州市土壤环境背景值分异规律研究

通过布点、采样和化验分析,研究了温州市东、中部地区约3 129 km2的滨海平原和丘陵地带以及乌岩岭自然保护区约25 km2的山地土壤环境背景值的分异规律。结果表明:pH、有机质、粒度等理化指标及重金属等元素和营养元素在温州市土类间有明显分异。所有元素背景值表现为从山地黄壤→红壤→水稻土→潮土→滨海盐土逐渐升高。同时这些元素存在土层纵向和土属之间的分异,而且在红壤土属中存在明显的迁移和累积特点。Pb、Ni、Cr、Hg、F在石英质砂岩、花岗岩和凝灰岩发育的红壤中明显累积,Zn、Cd、Mn明显迁移。     

水文分异条件下光养生物膜藻类对氮/磷的响应

光养生物膜是河流生态系统中重要的初级生产者,在河流生物地球化学循环中扮演重要角色。然而,河流水文状态及营养盐差异引起的生境异质性对光养生物膜藻类的影响未知。本研究在微型跑道池模拟流水(0.5 m/s)和静水(0 m/s)条件的基础上,通过设置不同浓度氮(1.51, 2.51和5.51 mg/L)、磷(0.1, 0.2和0.4 mg/L)及氮磷比(8, 16和32)条件培养野外采集的原位生物膜及人工基质,探究水文分异和营养变化对河流光养生物膜藻类物种组成及密度的影响。非度量多维排序(NMDS)分析结果表明,差异水文条件下,原位和建群生物膜藻类群落在梯度氮、磷和氮磷比环境中均呈现明显分离(PERMANOVA,P < 0.001),且建群生物膜中各分组藻类群落具有更明显的差异。双因素方差分析结果表明,生物膜中硅藻、蓝藻和绿藻的生长对营养盐与水文变化的响应并不一致,其中磷处理中,磷与流速单一及其交互效应显著影响了大多数藻类的生长及建群(P < 0.001);而氮处理中,氮与流速的交互效应仅对建群生物膜藻类影响显著(P < 0.001)。研究结果也发现,藻类在静水环境更有利于建群生物膜的形成,且静水-高营养盐环境更有利于蓝藻和绿藻的生长。这些结果说明,生物膜建群初期易受到水文扰动的影响,且水文分异和氮、磷影响了光养生物膜藻类的响应模式,研究为河流生态保护提供了新视角。     

CD36/Src/ERK pathway is involved in process of monocyte differentiation into macrophages

AIM To investigate the role of fatty acid translocase (FAT/CD36) on differentiation of monocytes to macrophages. METHODS Human monocyte THP-1 cells were treated with phorbol 12-myristate 13-acetate (PMA) at 0, 100 and 200 μg /L. Small interfering RNA (siRNA) targeting CD36 (siCD36) was employed to knock down the expression of CD36 in THP-1 cells. The CD36 over-expression (CD36OE) cell line was constructed by transfection with a recombinant lentivirus containing CD36 cDNA. Optical microscopy and crystal violet staining were used to detect the monocyte morphological changes and adhesion ability. The protein expression of CD36 was measured by flow cytometry and Western blot. The mRNA levels of CD36, CD11b and CD80 were detected by real-time PCR. The protein levels of extracellular signal-regulated kinase (ERK） and Src tyrosine kinase were determined by Western blot. RESULTS The cellular adhesiveness of THP-1 cells was elevated in the process of monocytes differentiation, and the expression of CD36 was increased in this process as well (P<0.01). siCD36 was transfected into the THP-1 cells (CD36i group) and the silencing efficiency was approximately 80%. The cell surface area and cellular adhesiveness were significantly decreased in CD36i group compared with scrambled siRNA (NCi) group (P<0.01). The mRNA levels of CD11b and CD80 were decreased in CD36i group compared with NCi group (P<0.01). The cell surface area and cellular adhesiveness were increased in CD36OE group compared with empty vector (vector) group (P<0.05). The mRNA levels of CD11b and CD80 were increased in CD36OE group compared with vector group (P<0.01). The phosphorylation levels of ERK and Src were decreased in CD36i group compared with NCi group (P<0.05). CONCLUSION CD36 promotes the differentiation of human monocyte THP-1 cells to macrophages by increasing the phosphorylation of Src and further activating ERK.