Large-scale generation of megakaryocytes from human embryonic stem cells using transgene-free and stepwise defined suspension culture conditions.
Large-scale generation of megakaryocytes from human embryonic stem cells using transgene-free and stepwise defined suspension culture conditions.
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使用无转基因和逐步确定的悬浮培养条件从人类胚胎干细胞大规模产生巨核细胞
DOI：
10.1111/cpr.13002
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发表时间：
2021-04
影响因子：
8.5
通讯作者：
Li Y
中科院分区：
文献类型：
--
作者：
Zhang B;Wu X;Zi G;He L;Wang S;Chen L;Fan Z;Nan X;Xi J;Yue W;Wang L;Wang L;Hao J;Pei X;Li Y
从人多能干细胞中离体工程化生产巨核细胞（MK）和血小板（PLT）是解决临床上供体捐献的PLT短缺和提供用于输血的诱导PLT的替代方法。然而，观察到低产量，并且从没有遗传修饰的人多能干细胞产生临床上适用的MK和PLT仍然需要改进。我们定义了一种最佳的，逐步的和完全无异种的培养方案，用于从人胚胎干细胞（hESC）生成MK。为了从hESC大规模产生MK，我们改进了单层诱导方式，通过使用特殊的聚苯乙烯CellSTACK培养室来定义MK的三维（3D）和球形分化系统。3D制造系统可以在连续培养的16 - 18天内从hESC有效地产生大量MK。经过三阶段有序诱导过程后，每个CellSTACK培养室平均可收集3.4 × 108个CD 41 + MK。通过3D诱导从hESC获得的MK显示出IL-8、血小板反应蛋白-1和MMP-9的显著分泌。在我们的培养体系中，来自hESC的诱导细胞被证明具有MK的特性以及形成proPLT和释放PLT的功能。此外，我们从临床级hESC系中产生了临床适用的MK，并证实了这些细胞的生物安全性。我们开发了一种简单的、逐步的、3D的和完全无异种/无饲养层/无转基因的诱导系统，用于从hESC产生MK。hESC衍生的MK显示具有典型的MK特征和PLT形成能力。本研究进一步增强了多能干细胞衍生的MK或PLT的临床应用。我们开发了一个简单的，逐步的，三维（3D）和完全无异种/无饲养层/无转基因诱导系统，用于从人胚胎干细胞（hESC）产生MK。hESC衍生的MK显示具有典型的MK特征和PLT形成能力。本研究进一步增强了多能干细胞衍生的MK或PLT的临床应用。
Ex vivo engineered production of megakaryocytes (MKs) and platelets (PLTs) from human pluripotent stem cells is an alternative approach to solve shortage of donor‐donated PLTs in clinics and to provide induced PLTs for transfusion. However, low production yields are observed and the generation of clinically applicable MKs and PLTs from human pluripotent stem cells without genetic modifications still needs to be improved. We defined an optimal, stepwise and completely xeno‐free culture protocol for the generation of MKs from human embryonic stem cells (hESCs). To generate MKs from hESCs on a large scale, we improved the monolayer induction manner to define three‐dimensional (3D) and sphere‐like differentiation systems for MKs by using a special polystyrene CellSTACK culture chamber. The 3D manufacturing system could efficiently generate large numbers of MKs from hESCs within 16‐18 days of continuous culturing. Each CellSTACK culture chamber could collect on an average 3.4 × 108 CD41+ MKs after a three‐stage orderly induction process. MKs obtained from hESCs via 3D induction showed significant secretion of IL‐8, thrombospondin‐1 and MMP9. The induced cells derived from hESCs in our culture system were shown to have the characteristics of MKs as well as the function to form proPLTs and release PLTs. Furthermore, we generated clinically applicable MKs from clinical‐grade hESC lines and confirmed the biosafety of these cells. We developed a simple, stepwise, 3D and completely xeno‐free/feeder‐free/transgene‐free induction system for the generation of MKs from hESCs. hESC‐derived MKs were shown to have typical MK characteristics and PLT formation ability. This study further enhances the clinical applications of MKs or PLTs derived from pluripotent stem cells. We developed a simple, stepwise, three‐dimensional (3D) and completely xeno‐free/feeder‐free/transgene‐free induction system for generation of MKs from human embryonic stem cells (hESCs). hESC‐derived MKs were shown to have the typical MK characteristics and PLT formation ability. This study further enhances the clinical application of MKs or PLTs derived from pluripotent stem cells.
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影响因子：
6
作者：
Guan X;Qin M;Zhang Y;Wang Y;Shen B;Ren Z;Ding X;Dai W;Jiang Y
通讯作者：
Jiang Y
影响因子：
5.3
作者：
Baigger A;Eicke D;Yuzefovych Y;Pogozhykh D;Blasczyk R;Figueiredo C
通讯作者：
Figueiredo C
影响因子：
21.3
作者：
Ditadi A;Sturgeon CM;Tober J;Awong G;Kennedy M;Yzaguirre AD;Azzola L;Ng ES;Stanley EG;French DL;Cheng X;Gadue P;Speck NA;Elefanty AG;Keller G
通讯作者：
Keller G
DOI：
10.1007/978-1-62703-478-4_3
发表时间：
2013-01-01
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Ni, Zhenya;Knorr, David A;Kaufman, Dan S
通讯作者：
Kaufman, Dan S
影响因子：
46.9
作者：
Ng, Elizabeth S.;Azzola, Lisa;Elefanty, Andrew G.
通讯作者：
Elefanty, Andrew G.