Soria B
Institute of Bioengineering and Department of Physiology, School of Medicine, Miguel Hernández University, Alicante, Spain.
Differentiation. 2001 Oct;68(4-5):205-19. doi: 10.1046/j.1432-0436.2001.680408.x.
Stem cell biology is a new field that holds promise for in-vitro mass production of pancreatic beta-cells, which are responsible for insulin synthesis, storage, and release. Lack or defect of insulin produces diabetes mellitus, a devastating disease suffered by 150 million people in the world. Transplantation of insulin-producing cells could be a cure for type 1 and some cases of type 2 diabetes, however this procedure is limited by the scarcity of material. Obtaining pancreatic beta-cells from embryonic stem cells would overcome this problem. We have derived insulin-producing cells from mouse embryonic stem cells by a 3-step in-vitro differentiation method consisting of directed differentiation, cell-lineage selection, and maturation. These insulin-producing cells normalize blood glucose when transplanted into streptozotocin-diabetic mice. Strategies to increase islet precursor cells from embryonic stem cells include the expression of relevant transcription factors (Pdx1, Ngn3, Isl-1, etc), together with the use of extracellular factors. Once a high enough proportion of islet precursors has been obtained there is a need for cell-lineage selection in order to purify the desired cell population. For this purpose, we designed a cell-trapping method based on a chimeric gene that fuses the human insulin gene regulatory region with the structural gene that confers resistance to neomycin. When incorporated into embryonic stem cells, this fusion gene will generate neomycin resistance in those cells that initiate the synthesis of insulin. Not only embryonic, but also adult stem cells are potential sources for insulin-containing cells. Duct cells from the adult pancreas are committed to differentiate into the four islet cell types; other possibilities may include nestin-positive cells from islets and adult pluripotent stem cells from other origins. Whilst the former are committed to be islet cells but have a reduced capacity to expand, the latter are more pluripotent and more expandable, but a longer pathway separates them from the insulin-producing stage. The aim of this review is to discuss the different strategies that may be followed to in-vitro differentiate pancreatic beta-cells from stem cells.
干细胞生物学是一个充满前景的新领域，有望实现体外大量生产胰腺β细胞，而胰腺β细胞负责胰岛素的合成、储存和释放。胰岛素的缺乏或缺陷会导致糖尿病，这是一种全球有1.5亿人遭受折磨的毁灭性疾病。移植产生胰岛素的细胞可能治愈1型糖尿病和某些2型糖尿病病例，然而这一过程受到材料稀缺的限制。从胚胎干细胞中获取胰腺β细胞将克服这一问题。我们通过一种三步体外分化方法从小鼠胚胎干细胞中获得了产生胰岛素的细胞，该方法包括定向分化、细胞谱系选择和成熟。将这些产生胰岛素的细胞移植到链脲佐菌素诱导的糖尿病小鼠体内后，可使血糖正常化。从胚胎干细胞中增加胰岛前体细胞的策略包括相关转录因子（Pdx1、Ngn3、Isl-1等）的表达，以及细胞外因子的使用。一旦获得足够比例的胰岛前体细胞，就需要进行细胞谱系选择以纯化所需的细胞群体。为此，我们设计了一种基于嵌合基因的细胞捕获方法，该嵌合基因将人胰岛素基因调控区与赋予新霉素抗性的结构基因融合。当整合到胚胎干细胞中时，这种融合基因将在开始合成胰岛素的细胞中产生新霉素抗性。不仅胚胎干细胞，成体干细胞也是含胰岛素细胞的潜在来源。成年胰腺的导管细胞可分化为四种胰岛细胞类型；其他可能性可能包括胰岛中巢蛋白阳性细胞和其他来源的成年多能干细胞。虽然前者注定会成为胰岛细胞，但扩增能力有限，而后者具有更强的多能性和更高的扩增能力，但它们与产生胰岛素阶段之间的路径更长。本综述的目的是讨论从干细胞体外分化胰腺β细胞可能采用的不同策略。
