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这是我翻译的一篇关于胚体培养的文章,分享下. R* B ?6 _: [6 R* r
Multiple mesodermal lineage differentiation of Apodemus sylvaticus embryonic stem cells in vitro
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Abstract$ y0 q' Q4 O/ L6 F: y) ^8 {
Background: Embryonic stem (ES) cells have attracted significant attention from researchers around the world 9 _& |4 Y" x! q; C
because of their ability to undergo indefinite self-renewal and produce derivatives from the three cell lineages, which & |( g6 Z( N6 d' r! e2 W
has enormous value in research and clinical applications. Until now, many ES cell lines of different mammals have been
4 G, n/ n. N5 s# U" vestablished and studied. In addition, recently, AS-ES1 cells derived from Apodemus sylvaticus were established and , [, w4 o0 I3 J
identified by our laboratory as a new mammalian ES cell line. Hence further research, in the application of AS-ES1 cells,
5 R! Z( Q0 y! j! F6 zis warranted.( m! q+ P0 `+ a4 J. }7 h1 o' f
Results: Herein we report the generation of multiple mesodermal AS-ES1 lineages via embryoid body (EB) formation 6 \+ n0 K6 L4 R) _# d! h T
by the hanging drop method and the addition of particular reagents and factors for induction at the stage of EB
' J) a% `4 P$ Jattachment. The AS-ES1 cells generated separately in vitro included: adipocytes, osteoblasts, chondrocytes and
- G: U4 H- u0 n% g% Q' I2 Zcardiomyocytes. Histochemical staining, immunofluorescent staining and RT-PCR were carried out to confirm the
" A2 k6 |8 z$ i, V" b8 m( ^; qformation of multiple mesodermal lineage cells.
) ~* w+ d0 q' o& @# iConclusions: The appropriate reagents and culture milieu used in mesodermal differentiation of mouse ES cells also
2 D) O9 i$ M- ^8 p# R9 h% K( Iguide the differentiation of in vitro AS-ES1 cells into distinct mesoderm-derived cells. This study provides a better
' ?7 x2 e$ G1 `$ j0 Y7 T8 C2 |understanding of the characteristics of AS-ES1 cells, a new species ES cell line and promotes the use of Apodemus ES
; ~- f3 H/ B- I! h+ Bcells as a complement to mouse ES cells in future studies.
" W' o! X K% D* d% w% S. k$ d摘要
. q# {% ?! w/ h* u* N. J, \1 g背景:胚胎干细胞(ES)的研究已经引起了全世界的重视,因为它们可以自我更新和由三个细胞系不断的增值产生新的细胞,在科研和临床有巨大的应用价值。到目前为止,很多哺乳动物胚胎干细胞系分别得到了建立和研究。此外,最近从林姬鼠得到的AS-ES1细胞作为一种新型的哺乳动物胚胎干细胞系在我们实验室成功建立。因此对于AS-ES1细胞的应用是值得进一步研究的。
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结果:此文中我们采用悬滴培养法获得胚体(EB)并在形成过程中加入特异性的诱导剂,获得多谱系的中胚层AS-ES1。AS-ES1细胞在体外分别分化为脂肪细胞,软骨细胞和成骨细胞,心肌细胞。免疫组织化学染色,荧光免疫染色和RT-PCR等对形成的多谱系中胚层细胞进行确认。
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: r2 u) T/ Q; ^. g9 g) F6 x' Q% f结论: 在老鼠的胚胎干细胞中胚层分化中使用的合适的试剂和培养基可以指导AS-ES1中胚层体外分化。本研究提供一个更好的了解AS-ES1细胞的特点, 将作为一种新的物种补充鼠科胚胎干细胞系。: D4 J! d- \) b6 n7 w
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