Zhang Shanshan, Morita Maiko, Wang Zhao, Ooehara Jun, Zhang Sen, Xie Miner, Bai Haitao, Yu Wenying, Wang Xiaofang, Dong Fang, Wang Jinhong, Ma Shihui, Yamazaki Satoshi, Ema Hideo
Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, China.
Project Division of Advanced Regenerative Medicine, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
Blood Sci. 2019 Sep 17;1(1):92-101. doi: 10.1097/BS9.0000000000000002. eCollection 2019 Aug.
Hematopoietic stem cells (HSCs) self-renew or differentiate through division. Cytokines are essential for inducing HSC division, but the optimal cytokine combination to control self-renewal of HSC in vitro remains unclear. In this study, we compared the effects of interleukin-12 (IL-12) and thrombopoietin (TPO) in combination with stem cell factor (SCF) on in vitro self-renewal of HSCs. Single-cell assays were used to overcome the heterogeneity issue of HSCs, and serum-free conditions were newly established to permit reproduction of data. In single-cell cultures, CD150CD48CD41CD34c-KitSca-1lineage HSCs divided significantly more slowly in the presence of SCF+IL-12 compared with cells in the presence of SCF+TPO. Serial transplantation of cells from bulk and clonal cultures revealed that TPO was more effective than IL-12 at supporting in vitro self-renewal of short-term (<6 months) HSCs, resulting in a monophasic reconstitution wave formation, whereas IL-12 was more effective than TPO at supporting the in vitro self-renewal of long-term (>6 months) HSCs, resulting in a biphasic reconstitution wave formation. The control of division rate in HSCs appeared to be crucial for preventing the loss of self-renewal potential from their in vitro culture.
造血干细胞（HSCs）通过分裂进行自我更新或分化。细胞因子对于诱导造血干细胞分裂至关重要，但体外控制造血干细胞自我更新的最佳细胞因子组合仍不清楚。在本研究中，我们比较了白细胞介素-12（IL-12）和血小板生成素（TPO）与干细胞因子（SCF）联合使用对造血干细胞体外自我更新的影响。采用单细胞分析来克服造血干细胞的异质性问题，并新建立了无血清条件以实现数据的重现。在单细胞培养中，与存在SCF+TPO的细胞相比，CD150CD48CD41CD34c-KitSca-1谱系造血干细胞在存在SCF+IL-12的情况下分裂明显更慢。来自批量和克隆培养细胞的连续移植显示，在支持短期（<6个月）造血干细胞的体外自我更新方面，TPO比IL-12更有效，导致单相重建波形成，而在支持长期（>6个月）造血干细胞的体外自我更新方面，IL-12比TPO更有效，导致双相重建波形成。控制造血干细胞的分裂速率对于防止其体外培养中自我更新潜能的丧失似乎至关重要。
