Lawrence Moyra, Shahsavari Arash, Bornelöv Susanne, Moreau Thomas, McDonald Rebecca, Vallance Thomas M, Kania Katarzyna, Paramor Maike, Baye James, Perrin Marion, Steindel Maike, Jimenez-Gomez Paula, Penfold Christopher, Mohorianu Irina, Ghevaert Cedric
Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, Cambridge Biomedical Campus, Puddicombe Way, Cambridge CB2 0AW, UK.
Department of Haematology and NHS Blood and Transplant, University of Cambridge, Cambridge, UK.
Sci Adv. 2022 Feb 18;8(7):eabj8618. doi: 10.1126/sciadv.abj8618. Epub 2022 Feb 16.
Platelet deficiency, known as thrombocytopenia, can cause hemorrhage and is treated with platelet transfusions. We developed a system for the production of platelet precursor cells, megakaryocytes, from pluripotent stem cells. These cultures can be maintained for >100 days, implying culture renewal by megakaryocyte progenitors (MKPs). However, it is unclear whether the MKP state in vitro mirrors the state in vivo, and MKPs cannot be purified using conventional surface markers. We performed single-cell RNA sequencing throughout in vitro differentiation and mapped each state to its equivalent in vivo. This enabled the identification of five surface markers that reproducibly purify MKPs, allowing us insight into their transcriptional and epigenetic profiles. Last, we performed culture optimization, increasing MKP production. Together, this study has mapped parallels between the MKP states in vivo and in vitro and allowed the purification of MKPs, accelerating the progress of in vitro-derived transfusion products toward the clinic.
血小板缺乏症，即血小板减少症，可导致出血，需通过输注血小板进行治疗。我们开发了一种从多能干细胞生产血小板前体细胞——巨核细胞的系统。这些培养物可维持100多天，这意味着巨核细胞祖细胞（MKP）能够实现培养更新。然而，目前尚不清楚体外的MKP状态是否反映体内状态，并且无法使用传统表面标志物纯化MKP。我们在整个体外分化过程中进行了单细胞RNA测序，并将每个状态与其体内等效状态进行映射。这使得能够鉴定出五种可重复纯化MKP的表面标志物，从而让我们深入了解它们的转录和表观遗传特征。最后，我们进行了培养优化，提高了MKP的产量。总之，本研究绘制了体内和体外MKP状态之间的相似之处，并实现了MKP纯化，加速了体外衍生的输血产品向临床应用的推进。
