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.     

春兰根状茎增殖与分化培养

以10种培养基进行了春兰根状茎增殖培养研究,结果表明,不同培养基对增殖率有明显影响,以White NAA 5 mg/L培养再转入1/2 MS NAA 5 mg/L培养基,可获得较高的平均生长速度,但获得的根状茎总量以2号培养基最多。春兰根状茎的最适增殖条件为:1/2 MS为基本培养基,不加或添加少量NAA,60 d为1个增殖周期;5种培养基分化培养结果表明,根状茎分化受BA浓度影响较大,当BA浓度达3 mg/L时,经45 d培养平均每g根状茎可分化出20个芽,每芽重约0.15 g。活性炭的存在会抑制分化。     

海岛棉愈伤组织诱导及分化的影响因素初探

本研究以新疆自育海岛棉品种新海16无菌苗下胚轴为外植体材料,进行愈伤组织诱导、分化及再生植株的初步研究,筛选出适合于海岛棉体细胞胚胎再生的培养体系。结果表明,最佳诱导愈伤组织及分化配方为MSB附加0.1mg/L2,4-D及0.05mg/LKT,分化率达到88%;于MSB1附加0.01mg/LKT及0.3mg/LNAA中对胚性愈伤组织进行增值培养;由MSB2附加0.1mg/LIBA中可分化出成熟体细胞胚胎及再生植株,分化率达73%。本研究通过体细胞胚胎发生途径获得了海岛棉再生植株,为海岛棉遗传转化体系的建立及分子育种奠定了基础。     

播期对洛麦21生长发育的影响

对洛麦21的生育规律进行了研究。结果表明:洛麦21的幼穗分化总体表现为前期慢、后期快,随着播期的推迟,幼穗分化的时间缩短。平均灌浆速率在1.453~1.697之间,在10月4日播种下平均灌浆速率最大,灌浆时间35~38 d,千粒重稳定在45 g以上。针对灌浆速率而言,以10月上旬适期播种对洛麦21最为有利。洛麦21主要依靠主茎和第1、2个一级分蘖成穗。随着播期的推迟,洛麦21的总分蘖数明显降低,成穗率提高。以10月14日播种的产量最高。     

野生与家养鹌鹑种群杂交后代遗传变异机制初探

不同种群间的杂交是丰富遗传变异最常见的手段,也是保护野生濒危物种常用的方法之一,研究杂交后群体遗传变异水平和机制对动物遗传资源保护、评价具有重要的意义。运用微卫星DNA标记对中国境内分布的野生日本鸣鹑与家鹌鹑群体及其杂交后代(F1、F2代)的遗传变异机制进行分析,结果表明:F1代鹌鹑群体每个座位平均检测出4.14个等位基因,群体多态信息含量和平均杂合度分别为0.5834和0.6265,具有比杂交亲本更高的遗传多样性水平。F2代遗传多样性有所降低,平均检测出3.57个等位基因,有4个等位基因在杂交过程中因遗传漂变而丢失;同时发现F1代、F2代与杂交亲本的遗传分化趋势不一致,F2代鹌鹑与野生日本鸣鹑遗传分化系数最大,为0.0712;而F1与之最小,仅为0.0275,表明鹌鹑杂交后代的遗传分化受分散过程的作用,主要受遗传漂变的影响。表明杂交可能是造成群体无定向分化及遗传多样性丧失的主要机制之一。     

文冠果花性状多尺度分析

针对文冠果新品种鉴定确立DUS性状指标体系与判别方法的需要,以白花、红花两种遗传类型的花性状为研究对象,选择两个地区作对比,从多尺度分析了9个花朵性状、9个花序性状的差异性和相关关系。性状差异性分析结果表明,花序性状比花朵性状稳定,白花花朵性状比红花花朵性状稳定;花性状的遗传变异大于在尺度上的变异;花性状的变异程度随着尺度的增大而降低,即由大到小的排序为单花尺度、花序尺度、植株尺度、区域尺度。性状相关性分析结果表明,白花性状间相关性大于红花的;花朵尺度性状间相关性大于花序尺度的;北京花朵尺度性状间相关性大于辽宁的,辽宁花序尺度性状间相关性大于北京的。首次建立了花冠面积与花瓣长、花瓣宽的二元指数估计模型;建立了花序质量与花序基径、花序长的一元指数估计模型,花序质量与花序基径、花序长和花序宽的三元指数估计模型,给出的模型可用于性状评价、选优、可塑性研究等,为文冠果种质资源鉴别、分类和多目标选育优良品系与品种鉴定提供依据。     

菊花胚性愈伤组织诱导及分化研究

以12个菊花品种为试材,采用三步培养法研究了胚性愈伤组织诱导、分化的特点。结果表明：胚性愈伤组织共有4种类型,其中黄绿色致密颗粒状胚性愈伤组织最好;‘玉人面’、‘铺地金’、‘紫妍’、‘杭白菊’间接体细胞胚发生的效果较好。