Chemically defined conditions for long-term maintenance of pancreatic progenitors derived from human induced pluripotent stem cells
Chemically defined conditions for long-term maintenance of pancreatic progenitors derived from human induced pluripotent stem cells
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DOI：
10.1038/s41598-018-36606-7
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发表时间：
2019-01-24
影响因子：
4.6
通讯作者：
Iwata, Hiroo
中科院分区：
文献类型：
--
作者：
Konagaya, Shuhei;Iwata, Hiroo
通过细胞移植来治疗I型糖尿病患者需要大量的分泌细胞，大约109个内分泌细胞。S 0X 9阳性胰腺上皮在胰腺发育的早期阶段广泛增殖。S 0X 9阳性胰腺上皮被认为是用于移植治疗的β细胞的可扩展细胞来源。在这项研究中，我们尝试使用化学成分确定的培养基在三维（3D）培养中扩增源自四种人iPSC系的胰腺祖细胞（PP：PDX 1（+）/SOX 9（+）），并检查衍生的PP分化为β样细胞的潜力。在含有表皮生长因子和R-spondin-1、CHIR 99021、成纤维细胞生长因子-7和SB 431542的化学成分确定的培养基中维持并有效增殖来自四种人iPSC系的PP。来源于一个iPSC系的PP可以在含有五种因子中的两种，表皮生长因子和R-spondin-1的化学成分确定的培养基中扩增超过10（4）倍。扩增的PP在冷冻保存后也是稳定的。冷冻和解冻后，PPs的增殖率没有下降。培养50天后获得的PP成功分化为胰岛素阳性β样细胞、胰高血糖素阳性α样细胞和生长抑素阳性δ样细胞。扩增PP的分化效率与未经扩增培养的PP的分化效率相似。
Large numbers of hormone-releasing cells, approximately 109 endocrine cells, are required to treat type I diabetes patients by cell transplantation. The SOX9-positive pancreatic epithelium proliferates extensively during the early stages of pancreatic development. SOX9-positive pancreatic epithelium is thought to be an expandable cell source of beta cells for transplantation therapy. In this study, we attempted to expand pancreatic progenitors (PPs: PDX1(+)/SOX9(+)) derived from four human iPSC lines in three-dimensional (3D) culture using a chemically defined medium and examined the potential of the derived PPs to differentiate into beta-like cells. PPs from four human iPSC lines were maintained and effectively proliferated in a chemically defined medium containing epidermal growth factor and R-spondin-1, CHIR99021, fibroblast growth factor-7, and SB431542. PPs derived from one iPSC line can be expanded by more than 10(4)-fold in chemically defined medium containing two of the fives, epidermal growth factor and R-spondin-1. The expanded PPs were also stable following cryopreservation. After freezing and thawing, the PPs proliferated without a decrease in the rate. PPs obtained after 50 days of culture successfully differentiated into insulin-positive beta-like cells, glucagon-positive alpha-like cells, and somatostatin-positive delta-like cells. The differentiation efficiency of expanded PPs was similar to that of PPs without expansion culture.