Application of stem-cell media to explant culture of human periosteum: An optimal approach for preparing osteogenic cell material.
Application of stem-cell media to explant culture of human periosteum: An optimal approach for preparing osteogenic cell material.
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DOI：
10.1177/2041731413509646
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发表时间：
2013
影响因子：
8.2
通讯作者：
Takagi R
中科院分区：
文献类型：
--
作者：
Uematsu K;Nagata M;Kawase T;Suzuki K;Takagi R
作为我们使用培养骨膜片进行骨再生的临床试验的一部分，我们在STK1和STK3两种干细胞培养基中制备了培养骨膜片。将人骨膜在1%人血清补充STK1中扩增28天，在1%人血清补充STK1中扩增14天，然后在1%人血清补充STK3中扩增14天（1%人血清补充STK1+3），或在10%胎牛血清补充Medium 199中扩增28天（对照组）。1%人血清添加STK1和1%人血清添加STK1+3培养的骨膜片直径和DNA含量显著高于对照组，且多层结构显著。在1%人血清中添加STK1+3后，成骨细胞标志物的信使RNA显著上调。在1%人血清补充STK1+3时，骨桥蛋白免疫阳性染色和矿化在培养的骨膜片的广泛区域都很明显。裸鼠皮下植入后，在1%人血清补充STK1+3中扩增，培养的骨膜片成骨活性最高。在1%的人血清中扩增STK1+3成功诱导培养的骨膜片生长，同时保留成骨潜能，随后的成骨诱导促进了均匀细胞材料的产生。
As part of our clinical tests on bone regeneration using cultured periosteal sheets, here, we prepared cultured periosteal sheets in two types of stem-cell culture media, STK1 and STK3. Human periosteum was expanded either in 1% human serum–supplemented STK1 for 28 days, in 1% human serum–supplemented STK1 for 14 days followed by 1% human serum–supplemented STK3 for 14 days (1% human serum–supplemented STK1+3), or in 10% fetal bovine serum–supplemented Medium 199 for 28 days (control). Cultured periosteal sheet diameter and DNA content were significantly higher, and the multilayer structure was prominent in 1% human serum–supplemented STK1 and 1% human serum–supplemented STK1+3. The messenger RNA of osteoblastic markers was significantly upregulated in 1% human serum–supplemented STK1+3. Osteopontin-immunopositive staining and mineralization were evident across a wide area of the cultured periosteal sheet in 1% human serum–supplemented STK1+3. Subcutaneous implantation in nude mice following expansion in 1% human serum–supplemented STK1+3 produced the highest cultured periosteal sheet osteogenic activity. Expansion in 1% human serum–supplemented STK1+3 successfully induced cultured periosteal sheet growth while retaining osteogenic potential, and subsequent osteoblastic induction promoted the production of homogeneous cell material.