Bioengineering the human bone marrow microenvironment in liquefied compartments: a promising approach for the recapitulation of osteovascular niches

Abstract

Recreating the biological complexity of living bone marrow (BM) in a single in vitro strategy has faced many challenges. Most bioengineered strategies propose the co-culture of BM cellular components entrapped in different matrices limiting their migration and self-organization capacity or in open scaffolds enabling their escaping. We propose a methodology for fabricating a "human bone marrow-in-a-liquefied-capsule" to overcome these challenges, embracing the most important BM components in a single platform. Since free dispersion of the cells within the BM is an essential feature to maintain their in vivo properties, this platform provides a liquefied environment for the encapsulated cells to move freely and self-organize. Inside liquefied capsules, an engineered endosteal niche (eEN) is co-cultured with human umbilical cord cells, including endothelial cells and hematopoietic stem and progenitor cells (HSPCs). Two different human-like BM niches were recreated under static and dynamic systems. Although the culture of the engineered BM capsules (eBMC) in these different environments did not change the structural and compositional features of the BM niches, the biophysical stimulation potentiated the cellular intercommunication and the biomolecules secretion, demonstrating an enhanced in vitro bio performance. Moreover, while the eBMC without HSPCs provided the secretion of hematopoietic supportive factors, the presence of these cells recapitulated more closely the biological complexity of the native BM niches. This functional eBMC approach is an innovative platform capable of investigating several components and interactions of BM niches and how they regulate BM homeostasis and hematopoiesis. STATEMENT OF SIGNIFICANCE: The recapitulation of the multifaceted bone marrow (BM) microenvironment under in vitro conditions has gained intensive recognition to understand the intrinsic complexity of the native BM. While conventional strategies do not recapitulate the BM osteovascular niches nor give the cellular components a free movement, we report for the first time the development of human bone marrow-in-a-liquefied-capsule to overcome such limitations. Our engineered BM capsules (eBMC) partially mimic the complex structure, composition, and spatial organization of the native osteovascular niches present in the BM. This strategy offers a platform with physiological relevance to exploit the niches' components/networks and how they regulate the hematopoiesis and the initiation/progression of various BM-related pathologies.