Wei Ya-Ming, Cao Qiong, Zhou Hua-You, Xia Rong, Lan Jun-Cai, Meng Fan-Yi, Bai Hai
Department of Blood Transfusion, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2005 Dec;13(6):1076-81.
Umbilical cord blood stem cell transplantation (CBSCT) has made significant progress in treatment of lethal congenital or malignant disorders. Both the incidence and severity of GVHD from CBSCT were lower than that from bone marrow and peripheral blood stem cell transplantation, particularly for adult patients, but these advantages were also associated with higher rates of relapse. The immune-mediated effect of natural killer and cytotoxic T cells against residual tumor cells were shown to prevent relapse and to induce remission after bone marrow transplantation. To explore possibility of ex vivo expansion of T, NK and CD34(+) cells from umbilical cord blood, cord blood was expanded ex vivo with different combinations of cytokines, T and NK cells proliferation and differentiation were observed. CB MNCs were separated in Ficoll-Isopaque column and cultured in IMDM for 14 days with different recombinant cytokines. Cultured cells were collected and analyzed for progenitor/stem cell immunophenotyping at day 0, 3, 7, and 14 by using flow cytometry. The results indicated that all test groups cultured with different combinations of SCF, IL-3, IL-6, IL-7, IL-2 showed significant expansion of UCB MNC, compared with the group without cytokines. All test groups showed expansion effects on CD34(+) cells, CD34(+) percentage went up from 1.6% in fresh CB to the highest 11.9% in group D (SCF + IL-3, IL-6, IL-2). The CD34(+) cells peak displayed at day 7 of culture in group A and D, while in other two groups B and C appeared at day 14 of culture. The expansion multiple of CD34(+) cells in all test groups at day 7 of culture were from 10 to 50. The average value of CD3(+) T cell in fresh UCB was 18.7 +/- 4.3%, the CD3(+) T cells decreased sharply in the medium without any interleukin, while obvious increase were observed in the other test groups containing different combinations of cytokines. The maximal expansion multiple of CD3(+) T cells reached 2 times of the fresh UCB level. CD56(+) cells amounted to 3.6 +/- 1.9% of fresh UCB, CD56(+) cell number increased significantly only in medium containing IL-2. It is concluded that T cells, NK cells as well as stem/progenitor cells can be expanded in the same time from CB-MNC with the combinations of cytokines.
脐带血干细胞移植（CBSCT）在治疗致命性先天性或恶性疾病方面取得了重大进展。CBSCT引发的移植物抗宿主病（GVHD）的发生率和严重程度均低于骨髓和外周血干细胞移植，尤其是对成年患者而言，但这些优势也伴随着更高的复发率。自然杀伤细胞和细胞毒性T细胞对残留肿瘤细胞的免疫介导作用已被证明可预防骨髓移植后的复发并诱导缓解。为了探索从脐带血中体外扩增T细胞、自然杀伤细胞（NK）和CD34(+)细胞的可能性，用不同细胞因子组合对脐带血进行体外扩增，并观察T细胞和NK细胞的增殖与分化情况。将脐血单个核细胞（CB MNCs）在Ficoll-Isopaque柱中分离，然后在IMDM中用不同的重组细胞因子培养14天。在第0、3、7和14天收集培养细胞，并用流式细胞术分析祖细胞/干细胞的免疫表型。结果表明，与无细胞因子组相比，所有用干细胞因子（SCF）、白细胞介素-3（IL-3）、白细胞介素-6（IL-6）、白细胞介素-7（IL-7）、白细胞介素-2（IL-2）不同组合培养的试验组中，脐血单个核细胞均有显著扩增。所有试验组对CD34(+)细胞均有扩增作用，CD34(+)百分比从新鲜脐血中的1.6%升至D组（SCF + IL-3、IL-6、IL-2）的最高11.9%。A组和D组在培养第7天出现CD34(+)细胞峰值，而其他两组B组和C组在培养第14天出现峰值。所有试验组在培养第7天CD34(+)细胞的扩增倍数为10至50。新鲜脐血中CD3(+) T细胞的平均值为18.7 +/- 4.3%，在无任何白细胞介素的培养基中CD3(+) T细胞急剧减少，而在含有不同细胞因子组合的其他试验组中则观察到明显增加。CD3(+) T细胞的最大扩增倍数达到新鲜脐血水平的2倍。CD56(+)细胞占新鲜脐血的3.6 +/- 1.9%，仅在含有IL-2的培养基中CD56(+)细胞数量显著增加。结论是，通过细胞因子组合可同时从脐血单个核细胞中扩增T细胞、NK细胞以及干细胞/祖细胞。
