Native extracellular matrix, synthesized ex vivo by bone marrow or adipose stromal cells, faithfully directs mesenchymal stem cell differentiation

Abstract

Mesenchymal stem cells (MSCs) are highly responsive to cues in the microenvironment (niche) that must be recapitulated ex vivo to study their authentic behavior. In this study, we hypothesized that native bone marrow (BM)- and adipose (AD)-derived extracellular matrices (ECM) were unique in their ability to control MSC behavior. To test this, we compared proliferation and differentiation of bone marrow (BM)-derived MSCs when maintained on native decellularized ECM produced by BM versus AD stromal cells (i.e. BM- versus AD-ECM). We found that both ECMs contained similar types of collagens but differed in the relative abundance of each. Type VI collagen was the most abundant (≈60% of the total collagen present), while type I was the next most abundant at ≈30%. These two types of collagen were found in nearly equal proportions in both ECMs. In contrast, type XII collagen was almost exclusively found in AD-ECM, while types IV and V were only found in BM-ECM. Physically and mechanically, BM-ECM was rougher and stiffer, but less adhesive, than AD-ECM. During 14 days in culture, both ECMs supported BM-MSC proliferation better than tissue culture plastic (TCP), although MSC-related surface marker expression remained relatively high on all three culture surfaces. BM-MSCs cultured in osteogenic (OS) differentiation media on BM-ECM displayed a significant increase in calcium deposition in the matrix, indicative of osteogenesis, while BM-MSCs cultured on AD-ECM in the presence of adipogenic (AP) differentiation media showed a significant increase in Oil Red O staining, indicative of adipogenesis. Further, culture on BM-ECM significantly increased BM-MSC-responsiveness to rhBMP-2 (an osteogenic inducer), while culture on AD-ECM enhanced responsiveness to rosiglitazone (an adipogenic inducer). These findings support our hypothesis and indicate that BM- and AD-ECMs retain unique elements, characteristic of their tissue-specific microenvironment (niche), which promote retention of MSC differentiation state (i.e. “stemness”) during expansion and direct cell response to lineage-specific inducers. This study provides a new paradigm for precisely controlling MSC fate to a desired cell lineage for tissue-specific cell-based therapies.

Development and Angiogenic Potential of Cell-Derived Microtissues Using Microcarrier-Template

“…Collagen types I, II, III, and IV expression was analyzed with immunofluorescence staining. All MT conditions deposited all collagen types and fibronectin, although Collagen type IV was qualitatively the most abundant of all analyzed proteins for all three MT conditions ( Figure 6 ) in line with Marinkovic et al findings [ 57 ]. In PLA and Cultispher ® S MTs, collagen type IV was localized mostly at their surface, resembling the basement membrane structure [ 58 , 59 ].…”

Extracellular Matrix Synthesis and Remodeling by Mesenchymal Stromal Cells Is Context-Sensitive

“…However, the same study showed that particularly TGFβ1 was lost during decellularization [23]. MSC-derived ECM also have been used successfully to improve MSC growth, differentiation, and angiogenic properties [20,[24][25][26] or to modulate immune responses [27,28]. In contrast to the current study, however, MSC-derived ECM had not been used to mimic health and disease so far.…”

The extracellular matrix of hematopoietic stem cell niches

“…Large amounts of this collagen are deposited in the bone matrix [211] , where it can be detected in compact and trabecular bone [212] . In BM, collagen type I is mainly expressed by osteoblasts [218] but also by bone marrow stromal cells [219] , [220] including MSCs [221] . Its synthesis is regulated by several factors and cytokines.…”