Abstract

A Microchamber-Based Instantaneous Synthesis of Humanised the Bone Marrow Micro-Tympanic and Stacks

Author(s): Arunodaya Bisht

There is no high-throughput microtissue platform for generating microbone in bone marrow. Here, we describe a method for the assembly of microtissue arrays from bone marrow stromal cells (BMSCs) in vitro and their maturation into bone marrow ossicles in vivo. Discs with arrays of 50 microwells were used to assemble microtissues of 3×10 5 BMSCs each on nylon mesh supports. Microtissues were cultured in chondrogenic induction medium and then in hypertrophic medium, which promotes endochondral ossification. They were then transplanted into NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice and reconstituted into bone marrow ossicles. Mice were transplanted with 10 5 CD34+ cells from human cord blood. Microbone contained more human CD45+ cells than mouse bone marrow, but almost no human CD34+ progenitor cells. Human hematopoietic progenitor cells cycle rapidly at an unphysiological rate in the mouse bone marrow and the decreased CD34+ cell content within the ossicles supports the hypothesis that the humanized niche better controls progenitor cell cycling. match. Microtissue assemblies within microwells connected by nylon membrane supports offer an elegant method for the fabrication and manipulation of microtissue assemblies with bone organ-like properties. More generally, this approach and platform will help bridge the gap between in vitro microtissue manipulation and in vivo microtissue transplantation.