Production of cloned pigs from salivary gland-derived progenitor cells.

Mayuko Kurome, Ryo Tomii, Satoshi Ueno, Katsumi Hiruma, Shirou Matsumoto, Kenji Okumura, Kimitoshi Nakamura, Mitsuhito Matsumoto, Yuji Kaji, Fumio Endo, Hiroshi Nagashima
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引用次数: 19

Abstract

To achieve tissue stem cell transplantation in clinical settings, translational studies using large animal models are essential to confirm the efficacy and safety of therapy. Therefore, with the ultimate objective of constructing a porcine model of stem cell transplantation in the present study we attempted to clone pigs using porcine salivary gland-derived progenitor cells (pSGPs) as nuclear donors. Normal chromosomal compositions of pSGPs were maintained after five to eight passages (73%, 41 of 56). Cell cycle was efficiently synchronized in G(0)/G(1) phase after 2 days of serum-starved culture (79%). Characteristics of multipotent pSGPs, that is, CD49f and intracellular laminin staining patterns, were unchanged after serum-starved culture. Developmental rate of blastocysts from embryos reconstructed using pSGPs as nuclear donors was significantly higher when compared to embryos reconstructed using fetal fibroblasts (27.7%, 38 of 137 vs. 12.8%, 17 of 138; p < 0.05). When a total of 615 reconstructed embryos were transplanted into four recipient gilts, all gilts became pregnant and produced 12 piglets. These findings suggest that pSGPs represent appropriate donor cells for porcine somatic cell nuclear transfer.

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唾液腺衍生祖细胞克隆猪的研究。
为了在临床环境中实现组织干细胞移植,使用大型动物模型的转化研究对于确认治疗的有效性和安全性至关重要。因此,本研究的最终目的是构建猪干细胞移植模型,我们尝试用猪唾液腺衍生祖细胞(pSGPs)作为核供体克隆猪。5 ~ 8传代后psgp的染色体组成保持正常(73%,41 / 56)。血清饥饿培养2天后,细胞周期有效同步于G(0)/G(1)期(79%)。多能pSGPs的特征,即CD49f和细胞内层粘连蛋白染色模式,在血清饥饿培养后没有变化。使用psgp作为核供体重建的胚胎囊胚发育率显著高于使用胎儿成纤维细胞重建的胚胎(27.7%,38 / 137;12.8%,17 / 138;P < 0.05)。将615个重构胚胎移植到4只受体后备母猪中,所有后备母猪都怀孕并生产了12头仔猪。这些发现表明psgp是猪体细胞核移植的合适供体细胞。
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