Giada Ostinelli, Marie-Frédérique Gauthier, Nathalie Vernoux, Emilie Bernier, Tristan Dubé, Simon Marceau, Stéfane Lebel, Marie-Ève Tremblay, André Tchernof
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Here we characterize the process of liposecretion, the cellular features of DFAT cells and the role of the cell cycle.</p><p><strong>Methods: </strong>Primary adipocytes and adipocyte-derived pluripotent cells (APC) were isolated from human adipose tissue and mature adipocytes were dedifferentiated in ceiling culture. The intracellular organization of DFAT and APC were compared using transmission electron microscopy (TEM), and the changes of intracellular lipid content over time were tracked with Oil Red O. Finally, we tested whether liposecretion is a cell cycle-dependent phenomenon by cultivating mature adipocytes in ceiling culture with or without four different inhibitors of the cell cycle (AraC, Irinotecan, Vincristine and RO-3306).</p><p><strong>Results: </strong>DFAT cells were enriched in intracellular lipids, which are stored in small lipid droplets. 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引用次数: 0
摘要
背景：去分化脂肪（DFAT）细胞由于其多能潜能已被应用于再生医学。根据文献，脂肪细胞去分化的过程以脂肪分泌为特征，导致成纤维细胞样的增殖细胞群，细胞周期相关基因的表达增加。许多途径都涉及到这一过程，但细胞周期在脂肪细胞去分化中的作用尚未被研究。在这里，我们描述脂肪分泌的过程，DFAT细胞的细胞特征和细胞周期的作用。方法：从人脂肪组织中分离原代脂肪细胞和脂肪细胞来源的多能细胞（APC），在天花板培养中去分化成熟脂肪细胞。利用透射电镜（TEM）比较了DFAT和APC的细胞内组织，并用Oil Red o跟踪了细胞内脂质含量随时间的变化。最后，我们通过使用或不使用四种不同的细胞周期抑制剂（AraC、伊立替康、Vincristine和RO-3306）在顶培养中培养成熟脂肪细胞，来测试脂肪分泌是否是一种细胞周期依赖现象。结果：DFAT细胞内脂质富集，以小脂滴形式储存。此外，脂肪分泌是成熟脂肪细胞去分化的特征，其特征是被膜包裹的大脂滴的快速分泌。这种现象似乎被细胞周期蛋白依赖性激酶1 （CDK1）抑制剂RO-3306的存在所阻碍。结论：两种人类脂肪组织库均经历体外去分化，但内脏脂肪组织DFAT细胞比皮下来源的DFAT细胞保留更多的脂质。脂分泌的特点是膜包裹的脂滴的快速排出。这种现象依赖于CDK1，可能依赖于整合素介导的细胞粘附的存在。
Exploring the role of cell cycle regulation in human mature adipocyte dedifferentiation.
Background: Dedifferentiated fat (DFAT) cells have been used in regenerative medicine due to their multipotent potential. According to the literature, the process of adipocyte dedifferentiation is characterized by liposecretion which results in a fibroblastlike, proliferating cell population, with increased expression of genes related to cell cycle. A number of pathways have been implicated in the process, but the role of the cell cycle in adipocyte dedifferentiation has yet to be investigated. Here we characterize the process of liposecretion, the cellular features of DFAT cells and the role of the cell cycle.
Methods: Primary adipocytes and adipocyte-derived pluripotent cells (APC) were isolated from human adipose tissue and mature adipocytes were dedifferentiated in ceiling culture. The intracellular organization of DFAT and APC were compared using transmission electron microscopy (TEM), and the changes of intracellular lipid content over time were tracked with Oil Red O. Finally, we tested whether liposecretion is a cell cycle-dependent phenomenon by cultivating mature adipocytes in ceiling culture with or without four different inhibitors of the cell cycle (AraC, Irinotecan, Vincristine and RO-3306).
Results: DFAT cells were enriched in intracellular lipids, which are stored in small lipid droplets. In addition, liposecretion, which characterizes mature adipocyte dedifferentiation, is characterized by the rapid secretion of a large lipid droplet that is coated by a membrane. This phenomenon seems to be hindered by the presence of cyclin dependent kinase 1 (CDK1) inhibitor RO-3306.
Conclusion: Both human adipose tissue depots undergo dedifferentiation in vitro, but visceral adipose tissue DFAT cells retain more lipids than subcutaneous-derived DFAT cells. Liposecretion is characterized by the rapid ejection of a membrane-wrapped lipid droplet. This phenomenon is dependent on CDK1 and likely relies on the presence of integrin-mediated cellular adherence.
期刊介绍：
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