Functional Analysis Of Tgf-beta Family Signaling During

为了预防和治疗艾滋病，了解胚胎造血过程中T淋巴细胞的细胞谱系是如何决定的机制是很重要的。BMP更多地规定了中胚层组织的腹侧命运。造血细胞来源于最腹侧的中胚层，因此，BMPs被认为在造血组织中T淋巴细胞谱系的确定中起着重要作用。我们验证这一假说的基本策略是利用整个胚胎培养系统培养含有BMPs或BMP受体等位基因的胚胎，然后在胚胎的感兴趣区域微量注射表达Cre重组酶的质粒。通过这种方法，我们可以实现区域特异性的基因破坏。为此，首先，我们建立了早在5.5天胚胎的培养方法。我们发现，添加75%大鼠血清的DMEM对E5.5胚胎的效果好于100%的大鼠血清，而100%的大鼠血清对E5.75胚胎的效果更好。我们正在优化条件，以实现Cre依赖的基因被Cre质粒破坏。通过这项研究获得的知识可能会让我们更好地理解原始的和确定的造血。

For prevention and cure of AIDS, it is important to know the mechanism of how cell lineage for T lymphocyte is determined during embryonic hematopoiesis. BMPs specify the fate of mesodermal tissues more ventral. Hematopoietic cells are derived from the most ventral mesoderm, therefore, BMPs are hypothesized to play important roles during specification of T lymphocyte lineage in the hematopoietic tissues. Our basic strategy to prove this hypothesis is to culture embryos that have floxed alleles for BMPs or BMP receptors using whole embryo culture system, and then to microinject Cre recombinase expressing plasmids in the interested region of embryos. Through this method, we could achieve region specific gene disruption. For this purpose, first, we established the culture method for the embryos as early as day 5.5 embryos. We found that DMEM supplemented with 75% rat serum worked better than 100 % rat serum for E5.5 embryos whereas 100 % rat serum works better for E5.75 embryos. We are underway to optimize the condition to achieve the Cre-dependent gene disruption by Cre plasmids. The knowledge that will be acquired through this research may give us better understanding for the primitive and definitive hematopoiesis.