Fan Xiubo, Bao Chunyu, Liu Yang, Liu Tianqing, Sun Xiangyu, Ge Dan, Ma Xuehu
Stem Cell and Tissue Engineering Lab of Dalian University of Technology, Dalian 116024, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2007 Aug;24(4):870-5.
Rotating wall vessel (RWV) was used for the ex-vivo expansion of umbilical cord blood stem cells to meet the requirement of clinical application in the aspect of quantity and quality of the stem cells. The mononuclear cells (MNCs) from umbilical cord blood were cultured in T-flasks for 24 h, and then inoculated in RWV to culture for 200 h. The nucleated-cell numbers, pH and osmolality of the culture medium were determined every 24 h. The CD34+ cells content was measured and CFU-GM culture was carried out at 144 h and 197 h. Nucleated cells (NC) and CD34+ cells had a 435.5 +/- 87.6 fold expansion and a 32.7 +/- 15.6 fold expansion respectively in 197 h, and CFU-GM (colony-forming unit-granulocyte/macrophage) cells had a 21.7 +/- 4.9 fold expansion. In the whole course of culture, the pH and osmolality of the medium in the RWV were kept in the optimal hematopoietic stem cells' expansion conditions. pH was kept from 7.2 to 7.4, and the osmolality was kept from 290 mmol/kg to 310 mmol/kg. Owing to its structural particularity, the RWV could ensure cells to grow in the suspension state, could simulate the micro-environment of umbilical cord blood, and thus could make the hematopoietic stem cells expand largely in the RWV in short time.
旋转壁式生物反应器（RWV）用于脐带血干细胞的体外扩增，以满足临床应用对干细胞数量和质量的要求。将脐带血中的单个核细胞（MNCs）在T型烧瓶中培养24小时，然后接种到RWV中培养200小时。每24小时测定一次培养基中的有核细胞数、pH值和渗透压。在144小时和197小时测量CD34+细胞含量并进行CFU-GM培养。有核细胞（NC）和CD34+细胞在197小时分别扩增了435.5±87.6倍和32.7±15.6倍，CFU-GM（集落形成单位-粒细胞/巨噬细胞）细胞扩增了21.7±4.9倍。在整个培养过程中，RWV中培养基的pH值和渗透压保持在造血干细胞扩增的最佳条件。pH值保持在7.2至7.4之间，渗透压保持在290 mmol/kg至310 mmol/kg之间。由于其结构特殊性，RWV可确保细胞在悬浮状态下生长，可模拟脐带血的微环境，从而可使造血干细胞在短时间内在RWV中大量扩增。
