A New High Throughput Screening Platform for Cell Encapsulation in Alginate Hydrogel Shows Improved Hepatocyte Functions by Mesenchymal Stromal Cells Co-encapsulation.

Valeria Iansante,Ragai R Mitry,Emer Fitzpatrick,Céline Filippi,Simon Walker,Charlotte A Lee,Raquel Fernandez-Dacosta,Fatma Masmoudi,Anil Dhawan

doi:10.3389/fmed.2018.00216

Valeria Iansante, Ragai R Mitry + Show 7 more

Open Access

https://doi.org/10.3389/fmed.2018.00216

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Abstract

Hepatocyte transplantation has emerged as an alternative to liver transplant for liver disease. Hepatocytes encapsulated in alginate microbeads have been proposed for the treatment of acute liver failure, as they are able to provide hepatic functions while the liver regenerates. Furthermore, they do not require immunosuppression, as the alginate protects the hepatocytes from the recipient's immune cells. Mesenchymal stromal cells are very attractive candidates for regenerative medicine, being able to differentiate into cells of the mesenchymal lineages and having extensive proliferative ability. When co-cultured with hepatocytes in two-dimensional cultures, they exert a trophic role, drastically improving hepatocytes survival and functions. In this study we aimed to (i) devise a high throughput system (HTS) to allow testing of a variety of different parameters for cell encapsulation and (ii) using this HTS, investigate whether mesenchymal stromal cells could have beneficial effects on the hepatocytes when co-encapsulated in alginate microbeads. Using our HTS platform, we observed some improvement of hepatocyte behavior with MSCs, subsequently confirmed in the low throughput analysis of cell function in alginate microbeads. Therefore, our study shows that mesenchymal stromal cells may be a good option to improve the function of hepatocytes microbeads. Furthermore, the platform developed may be used for HTS studies on cell encapsulation, in which several conditions (e.g., number of cells, combinations of cells, alginate modifications) could be easily compared at the same time.

Highlights

Liver transplantation represents the treatment of choice for patients with end-stage liver disease and liverbased metabolic disorders, in the last three decades human hepatocyte transplantation has emerged as a potential alternative [1,2,3,4,5]
We found that all the hepatic functions analyzed were significantly enhanced by Mesenchymal stromal cells (MSC) coencapsulation, confirming the results observed in alginate microdisks and further supporting the use of our high throughput system (HTS) platform as a reliable method for the initial pre-screening of encapsulation conditions
The total protein content (TP) measured on hepatocytes encapsulated in microbeads or microdisks was similar, showing a significant difference only at day 1, when cells were encapsulated in MD1 (TP = 32.6 ± 1.5 μg, p < 0.05), but not in MD2 (TP = 36.3 ± 2.4 μg), compared to MB (TP = 41.7 ± 3.2 μg) (Figure 2B)

Summary

Introduction

Liver transplantation represents the treatment of choice for patients with end-stage liver disease and liverbased metabolic disorders, in the last three decades human hepatocyte transplantation has emerged as a potential alternative [1,2,3,4,5]. Intraperitoneal transplantation of human hepatocytes encapsulated in alginate microbeads is an attractive option for the management of patients with ALF, as hepatocytes microbeads can provide hepatic functions while the patient’s own liver regenerates, with the advantage of not requiring any immunosuppression, as the alginate protects transplanted hepatocytes from the recipient’s immune cells [8, 9]. Theoretically readily available for cell transplantation, hepatocytes are not always suitable to this aim as their quality is often sub-optimal

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