Single Nucleus RNA Sequence (snRNAseq) Analysis of the Spectrum of Trophoblast Lineages Generated From Human Pluripotent Stem Cells in vitro.
Single Nucleus RNA Sequence (snRNAseq) Analysis of the Spectrum of Trophoblast Lineages Generated From Human Pluripotent Stem Cells in vitro.
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DOI：
10.3389/fcell.2021.695248
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发表时间：
2021
影响因子：
5.5
通讯作者：
Roberts RM
中科院分区：
文献类型：
--
作者：
Khan T;Seetharam AS;Zhou J;Bivens NJ;Schust DJ;Ezashi T;Tuteja G;Roberts RM
研究人类滋养细胞(TB)出现的一个模型是将多能干细胞暴露于骨形态发生蛋白4(BMP4)，其中存在激活素/TGFb抑制剂；A83-01和FGF2；PD173074(BAP)，它产生细胞滋养细胞、合体滋养细胞和与绒毛外滋养细胞相似的细胞。在此，H1人胚胎干细胞在两种氧气条件下(分别为20%和5%)暴露于BAP。在第8天，单核RNA测序用于转录分析，从而可以分析脆弱的合体结构以及更具弹性的单核细胞。在聚类分析之后，两个主要分组很明显，一个由五个(2，4，6，7，8)组成，第二个由三个(1，3，5)聚类组成，所有这些都显示出可识别的结核病标记。其中2例(2例和3例)与绒毛外滋养层细胞相似，2例(5例和6例)强烈携带合体滋养层细胞标志转录本，其余5例可能是不同类型的单核细胞滋养层细胞。我们认为，合体滋养层细胞内的两种核可能是由涉及两种不同前体细胞的融合事件引起的。O2条件下差异表达基因的数量在不同的簇之间有所不同，在5%O2培养的细胞中上调的基因数量在合体滋养层细胞簇6中最高。综上所述，BAP模型揭示了滋养层细胞谱系出现的出人意料的复杂图景，需要在时间过程研究中进一步解决。
One model to study the emergence of the human trophoblast (TB) has been the exposure of pluripotent stem cells to bone morphogenetic protein 4 (BMP4) in presence of inhibitors of ACTIVIN/TGFB; A83–01 and FGF2; PD173074 (BAP), which generates a mixture of cytotrophoblast, syncytiotrophoblast, and cells with similarities to extravillous trophoblast. Here, H1 human embryonic stem cells were BAP-exposed under two O2 conditions (20% and 5%, respectively). At day 8, single nuclei RNA sequencing was used for transcriptomics analysis, thereby allowing profiling of fragile syncytial structures as well as the more resilient mononucleated cells. Following cluster analysis, two major groupings, one comprised of five (2,4,6,7,8) and the second of three (1,3,5) clusters were evident, all of which displayed recognized TB markers. Of these, two (2 and 3) weakly resembled extravillous trophoblast, two (5 and 6) strongly carried the hallmark transcripts of syncytiotrophoblast, while the remaining five were likely different kinds of mononucleated cytotrophoblast. We suggest that the two populations of nuclei within syncytiotrophoblast may have arisen from fusion events involving two distinct species of precursor cells. The number of differentially expressed genes between O2 conditions varied among the clusters, and the number of genes upregulated in cells cultured under 5% O2 was highest in syncytiotrophoblast cluster 6. In summary, the BAP model reveals an unexpectedly complex picture of trophoblast lineage emergence that will need to be resolved further in time-course studies.
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影响因子：
4
作者：
Karvas, Rowan M.;McInturf, Samuel;Schulz, Laura C.
通讯作者：
Schulz, Laura C.
影响因子：
4.5
作者：
Ghazalpour A;Bennett B;Petyuk VA;Orozco L;Hagopian R;Mungrue IN;Farber CR;Sinsheimer J;Kang HM;Furlotte N;Park CC;Wen PZ;Brewer H;Weitz K;Camp DG 2nd;Pan C;Yordanova R;Neuhaus I;Tilford C;Siemers N;Gargalovic P;Eskin E;Kirchgessner T;Smith DJ;Smith RD;Lusis AJ
通讯作者：
Lusis AJ
影响因子：
7
作者：
Schneider, Valerie A.;Graves-Lindsay, Tina;Church, Deanna M.
通讯作者：
Church, Deanna M.
影响因子：
4.6
作者：
Jain, Ashish;Ezashi, Toshihiko;Tuteja, Geetu
通讯作者：
Tuteja, Geetu
影响因子：
14.9
作者：
Ortiz-Barahona A;Villar D;Pescador N;Amigo J;del Peso L
通讯作者：
del Peso L