Li Karen, Ooi Vincent E C, Chuen Carmen Ka Yee, Lam Audrey Carmen, Ooi Linda Shiou Mei, Zhang Xiao Bing, Tsang Kam Sze, Chiu Lawrence Chi Ming, Chan Kathy Yuen Yee, Li Chi Kong, Fok Tai Fai, Yuen Patrick Man Pan, Ng Pak Cheung
Li Ka Shing Institute of Health Sciences, Department of Paediatrics, The Chinese Uinversity of Hong Kong, Hong Kong, China.
Br J Haematol. 2008 Jan;140(1):90-8. doi: 10.1111/j.1365-2141.2007.06838.x. Epub 2007 Oct 1.
Ex vivo expansion of haematopoietic stem and progenitor cells in cytokine combinations is effective in promoting differentiation and proliferation of multilineage progenitor cells, but often results in reduction of self-renewable stem cells. This study investigated the effect of a mannose-binding lectin, NTL, purified from Narcissus tazetta var. chinensis, on prolonged maintenance and expansion of cord blood CD34+ cells. Our results showed that the presence of NTL or Flt-3 ligand (FL) significantly preserved a population of early stem/progenitor cells in a serum- and cytokine-free culture for 35 d. The effect of NTL on the ex vivo expansion of CD34+ cells in the presence of stem cell factor, thrombopoietin (TPO) and FL was also investigated. NTL-enhanced expansion of early progenitors (CD34+, CD34+CD38-, mixed colony-forming units and CFU-GEMM) and committed progenitor cells (granulocyte CFU, erythroid burst-forming units/CFU and megakayocyte CFU) after 8 and 12 d of culture. Six weeks after transplanting 12 d-expanded cells to non-obese diabetic severe combined immunodeficient mice, increased engraftment of human CD45+ cells was observed in the bone marrow of animals that received NTL-treated cells. The dual functions of NTL on long-term preservation and expansion of early stem/multilineage progenitor cells could be developed for applications in various cell therapy strategies, such as the clinical expansion of CD34+ cells for transplantation.
细胞因子组合中造血干细胞和祖细胞的体外扩增可有效促进多谱系祖细胞的分化和增殖,但通常会导致自我更新干细胞数量减少。本研究调查了从中国水仙中纯化得到的一种甘露糖结合凝集素NTL对脐带血CD34+细胞长期维持和扩增的影响。我们的结果表明,NTL或Flt-3配体(FL)的存在可在无血清和细胞因子的培养体系中显著维持一群早期干细胞/祖细胞达35天。同时还研究了在干细胞因子、血小板生成素(TPO)和FL存在的情况下NTL对CD34+细胞体外扩增的影响。培养8天和12天后,NTL增强了早期祖细胞(CD34+、CD34+CD38-、混合集落形成单位和CFU-GEMM)以及定向祖细胞(粒细胞CFU、红系爆式集落形成单位/ CFU和巨核细胞CFU)的扩增。将培养12天的细胞移植到非肥胖糖尿病严重联合免疫缺陷小鼠体内六周后,在接受NTL处理细胞的动物骨髓中观察到人类CD45+细胞的植入增加。NTL对早期干细胞/多谱系祖细胞的长期保存和扩增的双重作用可开发用于各种细胞治疗策略,如用于移植的CD34+细胞的临床扩增。