Establishment of cord blood collection method and hematopoietic stem cell expansion

(1)脐带血采集方法：用一次性注射器抽取脐带血，采集量为78.5 ± 35.8ml，细胞数为9.0 ± 5.1 × 10 ~（-8），单个核细胞数为3.8 ± 2.4 ± × 10 ~（-8）。灌流法细胞数为3.6 ± 3.5 × 10 ~（-8），单个核细胞数为1.2 ± 1.3 × 10 ~（-8）。无论是哪种方法，在采血量、采血细胞数上的个体差异都较大。两种方法之间没有任何统计学显著差异。脐带血库的灌注方法过于复杂。而且，也有一些情况下，脐带动脉导管无法虚拟插入。(2)脐带血细胞分离方法如考虑建立脐带血库，需去除红细胞，保存大量脐带血，因冷冻保存空间有限，需研究干细胞分离方法，有核细胞、MNC、CFU-GM的采集率分别为29.9 ± 10.0%，52.8 ± 16.8%，42.2 ± 30.4%，Percoll法。有核细胞、MNC和CFU-GM的采集率分别为66.9 ± 26.6%、63.2 ± 19.0%。9%，76.1 ± 53.1%。两组核细胞采集率差异有统计学意义（P < 0.01）。但两种方法对MNC和CFU-GM的采集率差异无统计学意义。Percoll法虽然含有的多核细胞较少，但与节省冷冻保存空间有关，但比较复杂。

(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.