Stromalized microreactor supports murine hematopoietic progenitor enrichment

Abstract

There is an emerging need to process, expand, and even genetically engineer hematopoietic stem and progenitor cells (HSPCs) prior to administration for blood reconstitution therapy. A closed-system and automated solution for ex vivo HSC processing can improve adoption and standardize processing techniques. Here, we report a recirculating flow bioreactor where HSCs are stabilized and enriched for short-term processing by indirect fibroblast feeder coculture. Mouse 3 T3 fibroblasts were seeded on the extraluminal membrane surface of a hollow fiber micro-bioreactor and were found to support HSPC cell number compared to unsupported BMCs. CFSE analysis indicates that 3 T3-support was essential for the enhanced intrinsic cell cycling of HSPCs. This enhanced support was specific to the HSPC population with little to no effect seen with the Lineage and Lineage cells. Together, these data suggest that stromal-seeded hollow fiber micro-reactors represent a platform to screening various conditions that support the expansion and bioprocessing of HSPCs ex vivo.

Influential factors for optimizing and strengthening mesenchymal stem cells and hematopoietic stem cells co-culture

“…Typically, a twodimensional layer of MSCs is used in the MSC and HSC co-culture to support CD34 + cell proliferation. Two-dimensional culture does not adequately mimic bone marrow microenvironmental conditions and reduces the opportunities to cultivate engraftable HSCs with long-term culture ability [22,47]. The threedimensional culture of MSCs in a co-culture system not only increases their positive effects on the HSCs proliferation but also preserves HSCs self-renewal and stemness potentialities [48].…”

“…In recent studies using of bioreactors, scaffolds, nanoparticles, and nano brils are diverse practical methods to improve expansion conditions of HSCs in vitro [22,23].…”

“…Co-culturing of MSCs with HSCs is commonly used for improving their proliferation. Numerous parameters such as cytokines, extracellular vehicles (EVs), scaffolding, three-dimensional culture, the source of MSCs, and hypoxia affect the co-culturing of these two cell types [17,22]. The culture conditions of these cells could be strengthened by manipulating these parameters.…”

Co-Culture of Mesenchymal Stem Cell Spheres with Hematopoietic Stem Cells Under Hypoxia: A Cost-Effective Method to Maintain Self-Renewal and Homing Marker Expression

“…Suspension cultivation of MSCs somehow primes them to show more supportive effects on HSCs [28,29]. Moreover, monolayer culturing of MSCs leads to diminished proliferation and secretory potentialities along with induced cellular senescence [30].…”