Fibronectin-Alginate microcapsules improve cell viability and protein secretion of encapsulated Factor IX-engineered human mesenchymal stromal cells

Abstract

Continuous delivery of proteins by engineered cells encapsu-lated in biocompatible polymeric microcapsules is of considerable therapeutic potential. However, this technology has not lived up to expectations due to inadequate cell--matrix interactions and subsequent cell death. In this study we hypoth-esize that the presence of fibronectin in an alginate matrix may enhance the viability and functionality of encapsulated human cord blood-derived mesenchymal stromal cells (MSCs) expressing the human Factor IX (FIX) gene. MSCs were encapsulated in alginate-PLL microcapsules containing 10, 100, or 500 μg/ml fibronectin to ameliorate cell survival. MSCs in microcapsules with 100 and 500 μg/ml fibronectin demonstrated improved cell viability and proliferation and higher FIX secretion compared to MSCs in non-supplemented microcapsules. In contrast, 10 μg/ml fibronectin did not significantly affect the viability and protein secretion from the encapsulated cells. Differentiation studies demonstrated osteogenic (but not chondrogenic or adipogenic) differentiation capability and efficient FIX secretion of the enclosed MSCs in the fibronectin-alginate suspension culture. Thus, the use of recombinant MSCs encapsulated in fibronectin-alginate microcapsules in basal or osteogenic cultures may be of practical use in the treatment of hemophilia B.

Gene therapy for hemophilia

“…Treatment of murine adipose tissue-derived stem/stromal cells (mADSCs) with LV can induce sustained hF.IX expression in vitro , though it remains to be seen whether these cells can engraft into a recipient and provide sustained correction (118). Similarly, menenchymal stem cells (MSCs) derived from human cord blood can be transduced with retroviral or LV vectors to produce F.IX in vitro and in vivo , though efforts are still underway to optimize the matrix for cell growth and F.IX production (119-122). …”

DoE Analysis of Approaches for Hydrogel Microbeads’ Preparation by Millifluidic Methods

“…The encapsulation of cells within HMBs plays a key role in cell transplant, isolating the cells from host tissue (providing immune protection) and maintaining the typical phenotype of cells, enhancing the production of biologically active molecules [ 4 ]. The possibility to immuno-isolate cells represents indeed a great advance for cell transplantation procedures, overcoming the constraints related to cell sources (i.e., allogeneic or xenogeneic cells), providing alternatives to autologous cells that are often scarcely available [ 5 ]. Hydrogel microbeads have been proposed to encapsulate different cell types, including islets of Langerhans, stem cells or inducing pluripotent stem cells, Hepatocytes, Chondrocytes, etc., to mention a few [ 6 , 7 ].…”

Assessment of the Behavior of Mesenchymal Stem Cells Immobilized in Biomimetic Alginate Microcapsules

“…Interestingly, our RGD-modified alginate matrices did not provide a suitable environment for chondrogenic differentiation of D1-MSCs. Previous studies showed that cell aggregates are needed to induce chondrogenic differentiation in MSCs in 3D cultures, , and the solid core of APA microcapsules does not facilitate that process. When the polymeric matrices have high restriction forces as in the case of our microcapsules, the cells normally proliferate based on the local cells and grow into small multispheroids hindering the chondrogenic differentiation .…”