Artificial in vitro blood production has been presented by recent literature as a necessary and achievable aim. In order to obtain the required hematopoietic stem cells (HSCs) proliferation and differentiation for mature blood cell production, studies have been conducted on using either cytokine-rich conditions or co-culturing with other cells. Alternatively, three-dimensional (3D) cell culture environments (such as tissue scaffolds) have been shown to affect cell morphology, proliferation and differentiation. Therefore, we investigated decellularized cancellous bones (DCBs), which provide 3D structure and natural extracellular matrix, as a scaffold for preserving and growing HSC niches in vitro. Additionally, we optimized a cell seeding method using mesenchymal stem cells as supporting cells. We discovered that, although adhering only to the top of DCBs when seeded at 37 °C, mesenchymal stem cells adhered to the inside of the scaffold at 4 °C, indicating that the seeding temperature is important to control the adherence ability of stem cells. This, in turn, was revealed to be important for HSC cell seeding on 3D extracellular matrix, and provides the required cell methodology to use DCBs as a great scaffold for blood cell production.

近来的文献已经提出了人工体外血液生产是必要且可实现的目标。为了获得成熟血细胞生产所需的造血干细胞（HSC）增殖和分化，已经在使用富含细胞因子的条件下或与其他细胞共培养的条件下进行了研究。或者，已显示三维（3D）细胞培养环境（例如组织支架）会影响细胞的形态，增殖和分化。因此，我们研究了脱细胞的松质骨（DCB），它具有3D结构和天然的细胞外基质，可作为体外保存和生长HSC壁ni的支架。此外，我们优化了使用间充质干细胞作为支持细胞的细胞接种方法。我们发现，虽然在37°C接种时仅粘附在DCB的顶部，但间充质干细胞在4°C时粘附在支架内部，这表明接种温度对于控制干细胞的粘附能力很重要。反过来，这被证明对于将HSC细胞接种在3D细胞外基质上很重要，并提供了使用DCB作为血细胞生产的重要支架所需的细胞方法。