Scaffolds composed of extracellular matrix (ECM) can assist tissue remodeling and repair following injury. The ECM is a complex biomaterial composed of proteins, glycoproteins, proteoglycans, and glycosaminoglycans, secreted by cells. The ECM contains fundamental biological cues that modulate cell behavior and serves as a structural scaffold for cell adhesion and growth. For clinical applications, where immune rejection is a constraint, ECM can be processed using decellularization methods intended to remove cells and donor antigens from tissue or organs, while preserving native biological cues essential for cell growth and differentiation. Recent studies show bioengineered organs composed by a combination of a diversity of materials and stem cells as a possibility of new therapeutic strategies to treat diseases that affect different tissues and organs, including the central nervous system (CNS). Nevertheless, the methodologies currently described for brain decellularization involve the use of several chemical reagents with many steps that ultimately limit the process of organ or tissue recellularization. Here, we describe for the first time a fast and straightforward method for complete decellularization of mice brain by the combination of rapid freezing and thawing following the use of only one detergent (Sodium dodecyl sulfate (SDS)). Our data show that using the protocol we describe here, the brain was entirely decellularized, while still maintaining ECM components that are essential for cell survival on the scaffold. Our results also show the cell-loading of the decellularized brain matrix with Neuro2a cells, which were identified by immunohistochemistry in their undifferentiated form. We conclude that this novel and simple method for brain decellularization can be used as a scaffold for cell-loading.

由细胞外基质 (ECM) 组成的支架可以帮助损伤后的组织重塑和修复。ECM 是一种复杂的生物材料，由细胞分泌的蛋白质、糖蛋白、蛋白聚糖和糖胺聚糖组成。ECM 包含调节细胞行为的基本生物学线索，并作为细胞粘附和生长的结构支架。对于免疫排斥受到限制的临床应用，ECM 可以使用脱细胞方法进行处理，旨在从组织或器官中去除细胞和供体抗原，同时保留对细胞生长和分化必不可少的天然生物线索。最近的研究表明，由多种材料和干细胞组合而成的生物工程器官可能是治疗影响不同组织和器官（包括中枢神经系统 (CNS)）疾病的新治疗策略的可能性。然而，目前描述的脑去细胞化方法涉及使用几种化学试剂，其中包含许多步骤，最终限制了器官或组织再细胞化的过程。在这里，我们首次描述了一种快速而直接的方法，通过仅使用一种洗涤剂（十二烷基硫酸钠 (SDS)）后快速冷冻和解冻的组合对小鼠大脑进行完全脱细胞。我们的数据表明，使用我们在这里描述的方案，大脑完全去细胞化，同时仍保持支架上细胞存活所必需的 ECM 成分。我们的结果还显示了去细胞化脑基质与 Neuro2a 细胞的细胞负载，这些细胞通过免疫组织化学鉴定为未分化形式。我们得出结论，这种新颖且简单的大脑去细胞化方法可用作细胞装载的支架。