建立人类造血类器官以评估造血损伤和再生效果。
Establishment of human hematopoietic organoids for evaluation of hematopoietic injury and regeneration effect.
机构信息
College of Chemistry & Materials Science, Hebei University, Hebei, Baoding, 071002, China.
Key Laboratory of Medicinal Chemistry and Molecular Diagnosis, Hebei University, Hebei, Baoding, 071002, China.
出版信息
Stem Cell Res Ther. 2024 May 4;15(1):133. doi: 10.1186/s13287-024-03743-y.
BACKGROUND
Human hematopoietic organoids have a wide application value for modeling human bone marrow diseases, such as acute hematopoietic radiation injury. However, the manufacturing of human hematopoietic organoids is an unaddressed challenge because of the complexity of hematopoietic tissues.
METHODS
To manufacture hematopoietic organoids, we obtained CD34 hematopoietic stem and progenitor cells (HSPCs) from human embryonic stem cells (hESCs) using stepwise induction and immunomagnetic bead-sorting. We then mixed these CD34 HSPCs with niche-related cells in Gelatin-methacryloyl (GelMA) to form a three-dimensional (3D) hematopoietic organoid. Additionally, we investigated the effects of radiation damage and response to granulocyte colony-stimulating factor (G-CSF) in hematopoietic organoids.
RESULTS
The GelMA hydrogel maintained the undifferentiated state of hESCs-derived HSPCs by reducing intracellular reactive oxygen species (ROS) levels. The established hematopoietic organoids in GelMA with niche-related cells were composed of HSPCs and multilineage blood cells and demonstrated the adherence of hematopoietic cells to niche cells. Notably, these hematopoietic organoids exhibited radiation-induced hematopoietic cell injury effect, including increased intracellular ROS levels, γ-H2AX positive cell percentages, and hematopoietic cell apoptosis percentages. Moreover, G-CSF supplementation in the culture medium significantly improved the survival of HSPCs and enhanced myeloid cell regeneration in these hematopoietic organoids after radiation.
CONCLUSIONS
These findings substantiate the successful manufacture of a preliminary 3D hematopoietic organoid from hESCs-derived HSPCs, which was utilized for modeling hematopoietic radiation injury and assessing the radiation-mitigating effects of G-CSF in vitro. Our study provides opportunities to further aid in the standard and scalable production of hematopoietic organoids for disease modeling and drug testing.
背景
人类造血类器官在模拟人类骨髓疾病方面具有广泛的应用价值，如急性造血辐射损伤。然而，由于造血组织的复杂性，制造人类造血类器官仍然是一个未解决的挑战。
方法
为了制造造血类器官，我们使用逐步诱导和免疫磁珠分选的方法从人类胚胎干细胞（hESCs）中获得 CD34 造血干细胞和祖细胞（HSPCs）。然后，我们将这些 CD34 HSPCs 与基质相关细胞混合在明胶甲基丙烯酰（GelMA）中形成三维（3D）造血类器官。此外，我们还研究了造血类器官中辐射损伤和对粒细胞集落刺激因子（G-CSF）的反应。
结果
GelMA 水凝胶通过降低细胞内活性氧（ROS）水平来维持 hESCs 衍生的 HSPCs 的未分化状态。在含有基质相关细胞的 GelMA 中建立的造血类器官由 HSPCs 和多谱系血细胞组成，并显示造血细胞与基质细胞的黏附。值得注意的是，这些造血类器官表现出辐射诱导的造血细胞损伤效应，包括细胞内 ROS 水平升高、γ-H2AX 阳性细胞百分比和造血细胞凋亡百分比增加。此外，在培养基中补充 G-CSF 可显著提高 HSPCs 的存活率，并增强这些造血类器官中辐射后的骨髓细胞再生。
结论
这些发现证实了成功地从 hESCs 衍生的 HSPCs 制造出初步的 3D 造血类器官，该类器官可用于模拟造血辐射损伤，并在体外评估 G-CSF 的辐射缓解作用。我们的研究为进一步促进造血类器官的标准化和规模化生产，以用于疾病建模和药物测试提供了机会。
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