Abstract
Mesenchymal stem/stromal cells (MSCs) exhibit undesired phenotypic changes during ex vivo expansion, limiting production of the large quantities of high quality primary MSCs needed for both basic research and cell therapies. Primary MSCs retain many desired MSC properties including proliferative capacity and differentiation potential when expanded on decellularized extracellular matrix (dECM) prepared from primary MSCs. However, the need to use low passage number primary MSCs (passage 3 or lower) to produce the dECM drastically limits the utility and impact of this technology. Here, we report that primary MSCs expanded on dECM prepared from high passage number (passage 25) human telomerase reverse transcriptase (hTERT) transduced immortal MSC cell lines also exhibit increased proliferation and osteogenic differentiation. Two hTERT-transduced placenta-derived MSC cell lines, CMSC29 and DMSC23 [derived from placental chorionic villi (CMSCs) and decidua basalis (DMSCs), respectively], were used to prepare dECM-coated substrates. These dECM substrates showed structural and biochemical differences. Primary DMSCs cultured on dECM-DMSC23 showed a three-fold increase in cell number after 14 days expansion in culture and increased osteogenic differentiation compared with controls. Primary CMSCs cultured on the dECM-DMSC23 exhibited a two-fold increase in cell number and increased osteogenic differentiation. We conclude that immortal MSC cell lines derived from different parts of the placenta produce dECM with varying abilities for supporting increased primary MSC expansion while maintaining important primary MSC properties. Additionally, this is the first demonstration of using high passage number cells to produce dECM that can promote primary MSC expansion, and this advancement greatly increases the feasibility and applicability of dECM-based technologies.
Highlights
Primary mesenchymal stem cells (MSCs) possess a long list of important properties required for regenerative medicine applications including high proliferative capacity, multi-lineage differentiation potential, ability to deposit extracellular matrix, and capacity to modulate the local immune environment
We recently reported the establishment and characterisation of immortalized DMSC and CMSC cell lines, referred to as DMSC23 and CMSC29 respectively, which were produced through human telomerase reverse transcriptase (hTERT) transduction
CMSC29 and DMSC23 cultures were decellularized according to previously published procedures in order to produce decellularized extracellular matrix (dECM)-coated cell culture surfaces [9]
Summary
Primary mesenchymal stem cells (MSCs) possess a long list of important properties required for regenerative medicine applications including high proliferative capacity, multi-lineage differentiation potential, ability to deposit extracellular matrix, and capacity to modulate the local immune environment. 105−106 primary MSCs can be isolated from bone marrow aspirate while the abovementioned therapies require a minimum of 108 highly functional primary MSCs per treatment. To achieve such a large number, the primary MSCs must undergo prolonged ex vivo expansion. By the time a sufficient quantity of cells is reached, a large percentage of the MSCs are “filler cells” that no longer possess the desired properties of primary MSCs [6,7,9,10] This technological shortcoming results in costly and inefficient production of primary MSCs and restricts current clinical use
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
