Isono Wataru, Kawasaki Tomoyuki, Ichida Justin K, Nagasaka Kazunori, Hiraike Osamu, Umezawa Akihiro, Akutsu Hidenori
Center for Regenerative Medicine, National Center for Child Health and Development, Setagaya, Tokyo, Japan.
Department of Obstetrics and Gynecology, Teikyo University School of Medicine, Itabashi, Tokyo, Japan.
Stem Cell Investig. 2023 Apr 19;10:10. doi: 10.21037/sci-2022-043. eCollection 2023.
Human pluripotent stem cells (hPSCs) such as embryonic stem cells (ESCs) and induced pluripotent stem cells (PSCs) have the capacity of self-renewal and multilineage differentiation . Conventional hPSCs, which are in a primed state, can produce various types of differentiated cells. However, the variability in their degree of pluripotency and differentiation propensities, which is influenced by the inductive methods and culture conditions, limit their availability. Therefore, PSCs in a naïve state are a promising source of PSCs.
We recently developed a culture system for naïve hPSCs using an inhibitor of the NOTCH signaling pathway and a histone H3 methyltransferase disruptor. This culture system requires feeder cells for stably maintaining the naïve hPSCs. We aimed to develop a culture system for hPSCs that could maintain pluripotency under feeder-free conditions.
We used two inhibitors to develop an alternative feeder-free culture system to obtain naïve hPSCs. The naïve cells underwent stable cellular proliferation and were positive for naïve stem cell markers; in addition, they could differentiate into the three germ layers. These feeder-free dome-shaped induced pluripotent stem cells (FFDS-iPSCs) have characteristics similar to that of naïve-like PSCs.
The naive hPSCs under feeder-free conditions could ensure supply of cells for various applications in regenerative medicine and disease modeling.
人类多能干细胞（hPSCs），如胚胎干细胞（ESCs）和诱导多能干细胞（iPSCs），具有自我更新和多向分化的能力。处于始发态的传统hPSCs能够产生各种类型的分化细胞。然而，其多能性程度和分化倾向的变异性受诱导方法和培养条件的影响，限制了它们的可用性。因此，处于原始态的PSCs是一种很有前景的PSCs来源。
我们最近利用NOTCH信号通路抑制剂和组蛋白H3甲基转移酶干扰剂开发了一种用于原始态hPSCs的培养系统。该培养系统需要饲养细胞来稳定维持原始态hPSCs。我们旨在开发一种能在无饲养层条件下维持多能性的hPSCs培养系统。
我们使用两种抑制剂开发了一种替代的无饲养层培养系统来获得原始态hPSCs。这些原始态细胞经历了稳定的细胞增殖，并且对原始态干细胞标志物呈阳性；此外，它们能够分化为三个胚层。这些无饲养层穹顶状诱导多能干细胞（FFDS-iPSCs）具有与原始态样PSCs相似的特征。
无饲养层条件下的原始态hPSCs可为再生医学和疾病建模的各种应用确保细胞供应。
