Unidad de Terapia Celular - Laboratorio de Patología Celular y Molecular, Centro de Medicina Regenerativa, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, Venezuela.
Int J Artif Organs. 2023 Jan;46(1):29-39. doi: 10.1177/03913988221136144. Epub 2022 Nov 9.
Under certain clinical and experimental conditions hematopoiesis occurs in other site than bone marrow (BM), such as the liver. Here, we develop a 3D organoid that mimics several components of the hematopoietic niche present during liver extramedullary hematopoiesis.
To evaluate the capacity of a 3D hematopoietic organoid (3D-HO) to function as a hematopoietic like-niche allowing for blood cell production outside of the BM.
The 3D-HO is constituted by liver sinusoidal endothelial cells (LSEC) as the stromal component, BM isolated from 5-FU treated mice (FU-BMCs), collagen microspheres and plasma clot as scaffolds. The ability of the 3D-HO to support the survival and functionality of FU-BMCs was investigated by using confocal microscopy, histology analysis, flow cytometry, and clonogenic assays.
After 15 and 30 days, post-ectopic implantation, histological studies of the 3D-HO showed the presence of cells with myeloid and lymphoid lineage morphology. Flow cytometry analysis of these cells showed the presence of cells expressing hematopoietic stem progenitor cells (HSPC) (Sca-1/c-Kit), myeloid (Gr-1) and lymphoid (B220 and CD19) markers. Clonogenic assays showed that cells from the 3D-HO formed hematopoietic colonies. Expression of the gene by cells from the 3D-HO, implanted for 30 days in female mice, indicated that male donor cells persist in this model of extramedullary hematopoiesis.
The 3D-HO constitutes an extramedullary hematopoietic-like niche which supports the survival and functionality of FU-BMCs. It may constitute an efficient model for investigating, and , those factors that control hematopoiesis outside BM.
在某些临床和实验条件下，造血发生在骨髓（BM）以外的部位，如肝脏。在这里，我们开发了一种 3D 类器官，模拟了肝外髓外造血过程中存在的造血龛的几个组成部分。
评估 3D 造血类器官（3D-HO）作为造血样龛的能力，使其能够在 BM 之外产生血细胞。
3D-HO 由肝窦内皮细胞（LSEC）作为基质成分、5-氟尿嘧啶（FU）处理的小鼠骨髓（FU-BMCs）、胶原微球和血浆凝块作为支架构成。通过共聚焦显微镜、组织学分析、流式细胞术和克隆形成实验研究 3D-HO 对 FU-BMCs 存活和功能的支持能力。
异位植入后 15 和 30 天，3D-HO 的组织学研究显示存在具有髓系和淋巴系形态的细胞。对这些细胞的流式细胞术分析显示存在表达造血干细胞祖细胞（HSPC）（Sca-1/c-Kit）、髓系（Gr-1）和淋巴系（B220 和 CD19）标志物的细胞。克隆形成实验表明，来自 3D-HO 的细胞形成了造血集落。植入雌性小鼠 30 天后，来自 3D-HO 的细胞表达 基因，表明雄性供体细胞在这种外髓外造血模型中持续存在。
3D-HO 构成了一个外髓外造血样龛，支持 FU-BMCs 的存活和功能。它可能构成一种有效的模型，用于研究、和那些控制 BM 外造血的因素。
