BetaLogics Venture, Janssen R & D LLC, Raritan, New Jersey, USA.
Stem Cells. 2013 Nov;31(11):2432-42. doi: 10.1002/stem.1489.
Human embryonic stem cells (hESCs) are considered a potential alternative to cadaveric islets as a source of transplantable cells for treating patients with diabetes. We previously described a differentiation protocol to generate pancreatic progenitor cells from hESCs, composed of mainly pancreatic endoderm (PDX1/NKX6.1-positive), endocrine precursors (NKX2.2/synaptophysin-positive, hormone/NKX6.1-negative), and polyhormonal cells (insulin/glucagon-positive, NKX6.1-negative). However, the relative contributions of NKX6.1-negative versus NKX6.1-positive cell fractions to the maturation of functional β-cells remained unclear. To address this question, we generated two distinct pancreatic progenitor cell populations using modified differentiation protocols. Prior to transplant, both populations contained a high proportion of PDX1-expressing cells (85%-90%) but were distinguished by their relatively high (80%) or low (~25%) expression of NKX6.1. NKX6.1-high and NKX6.1-low progenitor populations were transplanted subcutaneously within macroencapsulation devices into diabetic mice. Mice transplanted with NKX6.1-low cells remained hyperglycemic throughout the 5-month post-transplant period whereas diabetes was reversed in NKX6.1-high recipients within 3 months. Fasting human C-peptide levels were similar between groups throughout the study, but only NKX6.1-high grafts displayed robust meal-, glucose- and arginine-responsive insulin secretion as early as 3 months post-transplant. NKX6.1-low recipients displayed elevated fasting glucagon levels. Theracyte devices from both groups contained almost exclusively pancreatic endocrine tissue, but NKX6.1-high grafts contained a greater proportion of insulin-positive and somatostatin-positive cells, whereas NKX6.1-low grafts contained mainly glucagon-expressing cells. Insulin-positive cells in NKX6.1-high, but not NKX6.1-low grafts expressed nuclear MAFA. Collectively, this study demonstrates that a pancreatic endoderm-enriched population can mature into highly functional β-cells with only a minor contribution from the endocrine subpopulation.
人胚胎干细胞（hESCs）被认为是异体尸源胰岛的潜在替代物，可作为移植细胞来源用于治疗糖尿病患者。我们之前描述了一种从 hESC 分化生成胰腺祖细胞的方案，该方案生成的胰腺祖细胞主要为胰腺内胚层（PDX1/NKX6.1 阳性）、内分泌前体细胞（NKX2.2/synaptophysin 阳性、激素/NKX6.1 阴性）和多激素细胞（胰岛素/胰高血糖素阳性、NKX6.1 阴性）。然而，NKX6.1 阴性细胞与 NKX6.1 阳性细胞亚群对功能性β细胞成熟的相对贡献仍不清楚。为解决这个问题，我们使用改良的分化方案生成了两种不同的胰腺祖细胞群体。在移植前，两种群体均含有高比例的 PDX1 表达细胞（85%-90%），但 NKX6.1 的表达相对较高（80%）或较低（~25%）。NKX6.1 高和 NKX6.1 低祖细胞群体被分别移植到微囊化装置中，皮下移植到糖尿病小鼠体内。移植 NKX6.1 低细胞的小鼠在移植后 5 个月内持续高血糖，而 NKX6.1 高细胞的小鼠在 3 个月内糖尿病得到逆转。在整个研究过程中，各组间的空腹人 C 肽水平相似，但只有 NKX6.1 高细胞移植的小鼠在移植后 3 个月时表现出明显的餐时、葡萄糖和精氨酸反应性胰岛素分泌。NKX6.1 低细胞移植的小鼠表现出升高的空腹胰高血糖素水平。来自两组的 Theracyte 装置均几乎仅包含胰腺内分泌组织，但 NKX6.1 高细胞移植的小鼠中含有更多比例的胰岛素阳性和生长抑素阳性细胞，而 NKX6.1 低细胞移植的小鼠中主要含有胰高血糖素阳性细胞。NKX6.1 高细胞移植的胰岛素阳性细胞，但不是 NKX6.1 低细胞移植的胰岛素阳性细胞，表达核 MAFA。总之，这项研究表明，富含胰腺内胚层的细胞群可以成熟为具有高度功能性的β细胞，而仅由内分泌亚群产生少量贡献。
