Establishment of cord blood collection method and hematopoietic stem cell expansion

(1)脐带血采集方法。采用注射器法采集脐带血量、细胞数、单核细胞（MNC）数分别为78.5±35.8 ml、9.0±5.1 × 10e8、3.8±2.4±× 10e8（脐带血吸入一次性注射器）。灌注法细胞数、MNC数分别为3.6±3.5 × 10e8、1.2±1.3 × 10e8。两种方法在采血量、采血细胞数量等方面存在较大的个体差异。两种方法之间的差异无统计学意义。灌注法对于脐带血库来说过于复杂。而且，也有一些脐带动脉导管不能实际插入的情况。(2)脐带血细胞分离方法如果考虑引入脐带血银行，红细胞的移除是必要的，而且大量脐带血必须保存，干细胞分离方法由于有冷冻保存的空间而必须加以检验。Percoll法核细胞、MNC和CFU-GM的收集率分别为29.9±10.0%、52.8±16.8%和42.2±30.4%。核细胞、MNC和CFU-GM的收集率分别为66.9±26.6%、63.2±19.0%。HES法分别为（9%）、（76.1±53.1%）。核细胞收集率差异有统计学意义（p < 0.01）。但两种方法间MNC和CFU-GM的收集率差异无统计学意义。Percoll法虽然含有较少的多核细胞，但也需要节省冷冻保存的空间。

(1) The cord blood collection method.Collected cord blood quantity, cell number, mononuclear cell (MNC) number ware 78.5 ± 35.8 ml, 9.0 ± 5.1 x 10e8, 3.8 ± 2.4 ± × 10e8, respectively by Syringe method (cord blood was aspirated into a disposable syringe). And, cell number, MNC number ware, 3.6 ± 3.5 × 10e8, 1.2 ± 1.3 x 10e8, respectively by Perfusion method. The individual difference of which method as well as was big in the blood-collecting quantity blood, the number of the collected cells as well. There weren't any statistically significant differences between the methods. Perfusion method is too complicated for cord blood banking.. And, there were some cases which catheter the umbilical cord artery couldn't be virtually inserted into, too.(2) The cord blood cell separation methodIf the introduction of cord blood bank is thought about, red blood cells removal is necessary, and much cord blood must be kept, and stem cells separation method must be examined because of space for cryo-preservation.The collection rates of nuclear cells, MNC and CFU-GM ware 29.9 ± 10.0 %, 52.8 ± 16.8 %, 42.2 ± 30.4 %, respectively by Percoll method. And the collection rates of nuclear cells, MNC and CFU-GM ware 66.9 ± 26.6 %, 63.2 ± 19..9 %, 76.1 ± 53.1 %, respectively by HES method. There was statistically significant difference in the nuclear cell collection rate (p < 0.01). But, there were no statistically significant differences in the collection rates of MNC and CFU-GM between the methods. The Percoll method is a little complicated though there are few containing of the poly-nuclear cells and connected with saving space for cryo-preservation.