Roles of CD34+ cells and ALK5 signaling in the reconstruction of seminiferous tubule-like structures in 3-D re-aggregate culture of dissociated cells from neonatal mouse testes.
Roles of CD34+ cells and ALK5 signaling in the reconstruction of seminiferous tubule-like structures in 3-D re-aggregate culture of dissociated cells from neonatal mouse testes.
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CD34 细胞和 ALK5 信号传导在新生小鼠睾丸分离细胞 3-D 重新聚集培养物重建生精小管样结构中的作用。
DOI：
10.1371/journal.pone.0188705
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Shimamura K
中科院分区：
文献类型：
--
作者：
Abe SI;Abe K;Zhang J;Harada T;Mizumoto G;Oshikawa H;Akiyama H;Shimamura K
如果体外组织重建成功，可以提供一个精细而简单的系统来分析驱动形态发生和维持有序结构的潜在机制。最近，我们成功地重建生精索样和管样结构，使用3-D的分离睾丸细胞的再聚集培养。在睾丸形成中，内皮细胞从中肾迁移到胚胎性腺已被证明对睾丸索的发育至关重要，但内皮细胞如何促进睾丸索的形成仍不清楚。为了解释内皮细胞和管周细胞在重建索样和管样结构中的作用，我们研究了CD 34+内皮细胞和p75+细胞以及管周肌样细胞（PTMCs）在睾丸细胞三维重组培养中的行为。结果表明，这3种细胞都具有自身和相互间的再聚集能力，并在再聚集体中分离成3层，与体内的间质和管周组织非常相似。使用来自Sox 9-EGFP转基因小鼠的睾丸观察荧光支持细胞和其他非荧光类型的细胞的行为，显示在再聚集培养物中动态的细胞运动和分离。培养睾丸细胞剥夺间质和管周细胞导致畸形的结构，但重新添加它们恢复小管样结构。培养物中纯化的CD 34+细胞分化为p75+细胞和PTMC。这些结果表明，CD 34+细胞分化为p75+细胞，然后分化为PTMC。TGFβ信号传导抑制剂SB 431542和ALK 5i干扰了索样和管样结构的重建，后者损害了肾小管样和管周样结构的重建，以及CD 34+、p75+、PTMCs和Sertoli细胞的增殖及其运动和分化。这些结果表明，CD 34+细胞和通过ALK 5的信号传导在肾小管样、管周样和索样结构的形态发生中起关键作用。
Tissue reconstruction in vitro can provide, if successful, a refined and simple system to analyze the underlying mechanisms that drive the morphogenesis and maintain the ordered structure. We have recently succeeded in reconstruction of seminiferous cord-like and tubule-like structures using 3-D re-aggregate culture of dissociated testicular cells. In testis formation, endothelial cells that migrated from mesonephroi to embryonic gonads have been shown to be critical for development of testis cords, but how endothelial cells contribute to testis cord formation remains unknown. To decipher the roles of endothelial and peritubular cells in the reconstruction of cord-like and tubule-like structures, we investigated the behavior of CD34+ endothelial and p75+ cells, and peritubular myoid cells (PTMCs) in 3-D re-aggregate cultures of testicular cells. The results showed that these 3 types of cells had the capacity of re-aggregation on their own and with each other, and of segregation into 3 layers in a re-aggregate, which were very similar to interstitial and peritubular tissues in vivo. Observation of behaviors of fluorescent Sertoli cells and other non-fluorescent types of cells using testes from Sox9-EGFP transgenic mice showed dynamic cell movement and segregation in re-aggregate cultures. Cultures of testicular cells deprived of interstitial and peritubular cells resulted in dysmorphic structures, but re-addition of them restored tubule-like structures. Purified CD34+ cells in culture differentiated into p75+ cells and PTMCs. These results indicate that CD34+ cells differentiate into p75+ cells, which then differentiate into PTMCs. TGFβ signaling inhibitors, SB431542 and ALK5i, disturbed the reconstruction of cord-like and tubule-like structures, and the latter compromised re-construction of interstitial-like and peritubular-like structures, as well as the proliferation of CD34+, p75+, PTMCs, and Sertoli cells, and their movement and differentiation. These results indicate that CD34+ cells and signaling through ALK5 play pivotal roles in the morphogenesis of interstitial-like, peritubular-like and cord-like structures.
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影响因子：
2.5
作者：
Combes, Alexander N.;Lesieur, Emmanuelle;Harley, Vincent R.;Sinclair, Andrew H.;Little, Melissa H.;Wilhelm, Dagmar;Koopman, Peter
通讯作者：
Koopman, Peter
DOI：
10.1196/annals.1411.015
发表时间：
2007-01-01
期刊：
TESTICULAR CHROMOSOME STRUCTURE AND GENE EXPRESSION
影响因子：
--
作者：
Gassei, Kathrin;Schlatt, Stefan
通讯作者：
Schlatt, Stefan
影响因子：
3.7
作者：
Miles DC;Wakeling SI;Stringer JM;van den Bergen JA;Wilhelm D;Sinclair AH;Western PS
通讯作者：
Western PS
影响因子：
3.6
作者：
Mendis, Sirisha H. S.;Meachem, Sarah J.;Loveland, Kate L.
通讯作者：
Loveland, Kate L.
影响因子：
14
作者：
Alves-Lopes, Joao Pedro;Soder, Olle;Stukenborg, Jan-Bernd
通讯作者：
Stukenborg, Jan-Bernd