Chinese Journal of Tissue Engineering Research ›› 2017, Vol. 21 ›› Issue (34): 5487-8492.doi: 10.3969/j.issn.2095-4344.2017.34.013
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Received:
2017-10-30
Online:
2017-12-08
Published:
2018-01-04
About author:
Sun Hui-zhe, Master, Associate professor, Center for Medical Education Research, Shenyang Medical College, Shenyang 110034, Liaoning Province, China
Supported by:
CLC Number:
Sun Hui-zhe, Tian Wei, Zeng Liang, Qiu Jin-yun, Zhang Qian.
2.4 小肠黏膜下层海绵孔径大小的测量结果 冷冻干燥法制出的小肠黏膜下层海绵是多孔连通性分布的,质量浓度1%组支架的孔径为100-150 μm,质量浓度2%组的孔径为150-200 μm,其他两质量浓度组的孔径逐渐下降,此外,以质量浓度1%组支架各层之间的连接多数是较细的纤维般结构,导致整个支架内部结构较粗糙,利于细胞黏附(表2)。实验观察结果显示,在100 mmol/L EDC交联下1%质量分数的小肠黏膜下层海绵效果最好。 在50 mmol/L EDC交联下,质量浓度1%-2%组小肠黏膜下层海绵的孔径减少较小,2%-3%组孔径大小减小增大,3%-4%组孔径依旧减小,但无2%-3%组变化大。在100 mmol/L EDC交联下,质量浓度1%-2%组小肠黏膜下层海绵的孔径几乎无明显差异,2%-4%组孔径缩小趋势变大。在150 mmol/L EDC交联下,质量浓度1%-2%组肠黏膜下层海绵的孔径有细微增大,2%-3%组孔径明显缩小,3%-4%组孔径缩小程度减小(图4)。"
随着EDC交联浓度的升高,质量浓度1%组小肠黏膜下层海绵的孔径逐渐减少,2%组的孔径逐渐减少;3%组的孔径在交联浓度50-100 mmol/L时增大,在交联浓度100-150 mmol/L时明显缩小;4%组的孔径逐渐增大。 2.5 骨髓间充质干细胞培养结果 刚刚接种时,骨髓细胞悬液中的细胞大小不一,呈圆形,无法辨认其细胞核,2 d之后细胞开始贴壁,以多角形为主,还有梭形与三角形,悬浮血细胞在换液后逐渐被清除,纯化过程在2次换液后基本完成。细胞集落约在7 d后形成,没有接触抑制,大多数层次不清,形状不规则,2周后细胞则长满全层,见图5。 细胞鉴定:将1周的细胞传代,培在养瓶中放入多聚赖氨酸包被的载玻片,铺满后,将其取出,钙钴法碱性磷酸酶染色呈弱阳性,见图6。扫描电镜观察,小肠黏膜下层海绵上的骨髓基质干细胞多为星形和多角形,与光镜观察结果一致,见图7。"
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