King Natalie E, Courtney Jo-Maree, Brown Lachlan S, Fortune Alastair J, Blackburn Nicholas B, Fletcher Jessica L, Cashion Jake M, Talbot Jana, Pébay Alice, Hewitt Alex W, Morris Gary P, Young Kaylene M, Cook Anthony L, Sutherland Brad A
Tasmanian School of Medicine, College of Health and Medicine, University of Tasmania, Level 4, Medical Sciences Precinct, 17 Liverpool St, Hobart, TAS, 7000, Australia.
Menzies Institute for Medical Research, University of Tasmania, Hobart, Australia.
Stem Cell Res Ther. 2024 Mar 3;15(1):59. doi: 10.1186/s13287-024-03671-x.
Pericytes are multifunctional contractile cells that reside on capillaries. Pericytes are critical regulators of cerebral blood flow and blood-brain barrier function, and pericyte dysfunction may contribute to the pathophysiology of human neurological diseases including Alzheimers disease, multiple sclerosis, and stroke. Induced pluripotent stem cell (iPSC)-derived pericytes (iPericytes) are a promising tool for vascular research. However, it is unclear how iPericytes functionally compare to primary human brain vascular pericytes (HBVPs).
We differentiated iPSCs into iPericytes of either the mesoderm or neural crest lineage using established protocols. We compared iPericyte and HBVP morphologies, quantified gene expression by qPCR and bulk RNA sequencing, and visualised pericyte protein markers by immunocytochemistry. To determine whether the gene expression of neural crest iPericytes, mesoderm iPericytes or HBVPs correlated with their functional characteristics in vitro, we quantified EdU incorporation following exposure to the key pericyte mitogen, platelet derived growth factor (PDGF)-BB and, contraction and relaxation in response to the vasoconstrictor endothelin-1 or vasodilator adenosine, respectively.
iPericytes were morphologically similar to HBVPs and expressed canonical pericyte markers. However, iPericytes had 1864 differentially expressed genes compared to HBVPs, while there were 797 genes differentially expressed between neural crest and mesoderm iPericytes. Consistent with the ability of HBVPs to respond to PDGF-BB signalling, PDGF-BB enhanced and a PDGF receptor-beta inhibitor impaired iPericyte proliferation. Administration of endothelin-1 led to iPericyte contraction and adenosine led to iPericyte relaxation, of a magnitude similar to the response evoked in HBVPs. We determined that neural crest iPericytes were less susceptible to PDGFR beta inhibition, but responded most robustly to vasoconstrictive mediators.
iPericytes express pericyte-associated genes and proteins and, exhibit an appropriate physiological response upon exposure to a key endogenous mitogen or vasoactive mediators. Therefore, the generation of functional iPericytes would be suitable for use in future investigations exploring pericyte function or dysfunction in neurological diseases.
周细胞是位于毛细血管上的多功能收缩细胞。周细胞是脑血流和血脑屏障功能的关键调节因子，周细胞功能障碍可能导致包括阿尔茨海默病、多发性硬化症和中风在内的人类神经疾病的病理生理过程。诱导多能干细胞（iPSC）衍生的周细胞（i周细胞）是血管研究的一种有前景的工具。然而，尚不清楚i周细胞在功能上与原代人脑血管周细胞（HBVP）相比如何。
我们使用既定方案将iPSC分化为中胚层或神经嵴谱系的i周细胞。我们比较了i周细胞和HBVP的形态，通过qPCR和批量RNA测序对基因表达进行定量，并通过免疫细胞化学观察周细胞蛋白标志物。为了确定神经嵴i周细胞、中胚层i周细胞或HBVP的基因表达是否与其体外功能特征相关，我们在暴露于关键的周细胞促有丝分裂原血小板衍生生长因子（PDGF）-BB后对EdU掺入进行定量，并分别观察对血管收缩剂内皮素-1或血管舒张剂腺苷的收缩和舒张反应。
i周细胞在形态上与HBVP相似，并表达典型的周细胞标志物。然而，与HBVP相比，i周细胞有1864个差异表达基因，而神经嵴和中胚层i周细胞之间有797个基因差异表达。与HBVP对PDGF-BB信号作出反应的能力一致，PDGF-BB增强了i周细胞的增殖，而PDGF受体-β抑制剂则损害了i周细胞的增殖。给予内皮素-1导致i周细胞收缩，给予腺苷导致i周细胞舒张，其幅度与HBVP中诱发的反应相似。我们确定神经嵴i周细胞对PDGFRβ抑制的敏感性较低，但对血管收缩介质的反应最为强烈。
i周细胞表达与周细胞相关的基因和蛋白质，并在暴露于关键的内源性促有丝分裂原或血管活性介质时表现出适当的生理反应。因此，功能性i周细胞的产生将适用于未来探索神经疾病中周细胞功能或功能障碍的研究。
