小鼠早期胚胎在6℃不同贮液中的保存研究

低温能抑制哺乳动物植入前胚胎的代谢和发育，从而可使胚胎在体外进行短期贮存。本文利用三种不同贮存液：Ｗｈｉｔｔｅｎ′ｓ、ＰＢＳ和ＣＭＲＬ－１０６６对小鼠２－和８－细胞胚胎进行贮存研究。结果表明，贮于上述三种贮液中的小鼠２－细胞胚胎，在６℃贮２４小时后，经３７℃培养，成活率分别为４２．８％、０％和０％；贮７２小时后的８－细胞胚胎的成活率分别为４３．３％、２０．０％和０％。还证实，培养液中丙酮酸钠和乳酸钠的存在对小鼠胚胎的体外发育是至关重要的。     

中华绒螯蟹卵巢和胚胎发育期脂类在卵黄物质中存在的形态及其变化

脂类在中华绒螯蟹成熟卵子中以两种形态存在,脂肪滴和卵黄体,前者为中性脂,卵黄体中的脂类主要是磷脂。Ｌ和Ｙ几乎充满足整个成熟卵子,且交互分布,开始它们间以许多致密小颗粒间,直到两者的分布无界限分割。     

粘皮鲻虎鱼胚胎及仔鱼的发育

粘皮鲻鰕虎鱼(Mugilogobius myxodermus)的卵产出时为球型,直径0.40~0.60mm,具黏性,卵颜色淡黄,半透明,油球多且数量不固定。受精5~10min后吸水膨胀成椭圆形。水温16~18℃的条件下,胚胎发育共需138h。初孵仔鱼体长2.30~3.10mm,体高0.35~0.40mm,总肌节29~30。出膜30min~24h内鳔充气,从孵出到各鳍分化完全约需要40d。     

Identification of a novel population of human cord blood cells with hema-topoietic and chondrocytic potential

With the exception of mature erythrocytes, cells within the human hematopoietic system are characterized by the cell surface expression of the pan-leukocyte receptor CD45. Here, we identify a novel subset among mononuclear cord blood cells depleted of lineage commitment markers (Lin-) that are devoid of CD45 expression. Surprisingly, functional examination of Lin-CD45- ceils also lacking cell surface CD34 revealed they were capable of multipotential hematopoietic progenitor capacity. Co-culture with mouse embryonic limb bud cells demonstrated that Lin^-CD45^-CD34^- cells were capable of contributing to cartilage nodules and differentiating into human chondrocytes. BMP-4, a mesodermal factor known to promote chondrogenesis, significantly augmented Lin^-CD45^-CD34^-differentiation into chondrocytes. Moreover, unlike CD34~ human hematopoietic stem cells, Lin^-CD45^-CD34^- cells were unable to proliferate or survive in liquid cultures, whereas single Lin^-CD45^-CD34^- cells were able to chimerize the inner cell mass (1CM) of murine blastocysts and proliferate in this embryonic environment. Our study identifies a novel population of Lin-CD45-CD34^- cells capable of commitment into both hematopoietic and chondrocytic lineages, suggesting that human cord blood may provide a more ubiquitous source of tissue with broader developmental potential than previously appreciated.     

低氧对胚胎干细胞增殖的影响

目的:观察间歇性低氧和持续性低氧对体外培养的胚胎干细胞(ES细胞)增殖的影响.方法:利用细胞记数法和BrdU (5-溴脱氧尿苷)掺入的流式细胞分析检测细胞增殖,并用RT -PCR的方法检测低氧诱导因子(HIF-1a)的表达变化.结果:①将ES细胞分别放在低氧(3%～10% O2)和常氧(20% O2)的环境中培养24 h后,在低氧环境中培养的ES细胞数较常氧组明显减少;②将ES细胞分别给予间歇性低氧刺激(3%～10% O2),每天10 min,连续4 d后,发现3%低氧组较常氧对照组的细胞增殖明显升高.③用RT-PCR方法观察HIF-1a的表达与细胞增殖的关系,发现在常氧环境中培养的ES即有HIF-1a的表达,ES细胞在持续低氧24 h或间歇性低氧(3%～10% O2)刺激4 d后对HIF-1a的表达均无明显影响.结论:间歇性低氧(3% O2)可明显促进体外培养的ES细胞增殖,而持续性低氧抑制ES细胞增殖,间歇性低氧(3% O2)刺激促进ES细胞增殖的机制尚有待于进一步的研究.     

A new trick to tune down TGFβ signal

Signal transduction in early embryogenesis needs to be properly controlled. A new player involved in tuning down TGF β signaling has now been identified - new evidence that multi-layer control of signaling is essential in vivo.     

Delay of ZGA initiation occurred in 2-cell blocked mouse embryos

One-cell mouse embryos from KM strain and B6C3F1 strain were cultured in M16 medium, in which2-cell block generally occurs. Embryos of KM strain exhibited 2-cell block, whereas B6C3F1 embryos,which are regarded as a nonblocking strain, proceeded to the 4-cell stage in our culture condition. It is oftenassumed that the block of early development is due to the failure of zygotic gene activation (ZGA) in culturedembryos. In this study we examined protein synthesis patterns by two-dimensional gel electrophoresis of[35^S] methionine radiolabeled 2-cell embryos. Embryos from the blocking strain and the nonblocking strainwere compared in their development both in vitro and in vivo. The detection of TRC expression, a markerof ZGA, at 42 h post hCG in KM embryos developed in vitro suggested that ZGA was also initiated even inthe 2-cell arrested embryos. Nevertheless, a significant delay of ZGA was observed in KM strain as comparedwith normally developed B6C3F1 embryos. At the very beginning of major ZGA as early as 36 h post hCG,TRC has already been expressed in B6C3F1 embryos developed in vitro and KM embryos developed in vivo.But for 2-cell blocked KM embryos, TRC was still not detectable even at 38 h post hCG. These evidences suggest that 2-cell-blocked embryos do initiate ZGA, and that 2-cell block phenomenon is due not to the disability in initiating ZGA, but to a delay of ZGA.     

Overexpression of S-adenosylmethionine decarboxylase.（SAMDC） in Xeno-pus embryos activates maternal program of apoptosm as a “fail-safe”mechanism of early embryogenesis

In Xenopus, injection of S-adenosylmethionine decarboxylase (SAMDC) mRNA into fertilized eggs or 2-cell stage embryos induces massive cell dissociation and embryo-lysis at the early gastrula stage due toactivation of the maternal program of apoptosis. We injected SAMDC mRNA into only one of the animalside blastomeres of embryos at different stages of cleavage, and examined the timing of the onset of theapoptotic reaction. In the injection at 4-and 8-cell stages, a considerable number of embryos developed intotadpoles and in the injection at 16-and 32-cell stages, all the embryos became tadpoles, although tadpolesobtained were sometimes abnormal. However, using GFP as a lineage tracer, we found that descendant cellsof the blastomere injected with SAMDC mRNA at 8-to 32-cell stages are confined within the blastocoel atthe early gastrula stage and undergo apoptotic cell death within the blastocoel, in spite of the continued development of the injected embryos. These results indicate that cells overexpressed with SAMDC undergo apoptotic cell death consistently at the early gastrula stage, irrespective of the timing of the mRNA injection.We assume that apoptosis is executed in Xenopus early gastrulae as a “fall-safe“ mechanism to eliminate physiologically-severely damaged cells to save the rest of the embryo.     

人类胚胎对植入的调控

胚胎植入是一个十分复杂的过程,需要胚泡和子宫内膜之间的协同作用。植入过程中,胚泡和内膜之间的通讯和相互作用仍是生殖医学领域中尚未解决的问题。最近的研究取得了一些进展,发现在人类胚胎植入的定位和粘会过程中,胚胎能对子宫内膜上皮的分子,如趋化因子,粘附分子,抗粘附分子和瘦素进行旁分泌调控。     

冬小麦原生质体培养的胚状体直接发生

冬小麦品种“京花一号”胚性愈伤组织在改良的N₆培养基(NBD培养基)上继代得到易碎型胚性愈伤组织,转入改良MS液体培养基(MSDL培养基)后得到胚性悬浮系,分离的原生质体在改良的MS培养基(MSDP培养基)上培养,再生细胞直接产生体细胞胚胎,并再生出完整植株。体细胞胚胎形成过程与小麦合子胚的形成过程十分相似。