Enabling a robust scalable manufacturing process for therapeutic exosomes through oncogenic immortalization of human ESC-derived MSCs

Journal of Translational Medicine - 2011
Tian Sheng Chen1, Fatih Arslan2, Yajie Yin1, S. G. Tan1, Ruenn Chai Lai3, Andre Choo4, Jayanthi Padmanabhan4, Chuen Neng Lee5, Dominique P.V. de Kleijn2, Sai Kiang Lim5
1Institute of Medical Biology, A*STAR, 8A Biomedical Grove, 138648, Singapore
2Laboratory of Experimental Cardiology, University Medical Centre Utrecht, Heidelberglaan 100, 3584 CX, Utrecht, the Netherlands
3National University of Singapore, Graduate School for Integrative Sciences and Engineering, 28 Medical Drive, 117456, Singapore
4Bioprocessing Technology Institute, A*STAR, 20 Biopolis Way, 138671, Singapore
5Department of Surgery, YLL School of Medicine, NUS, 5 Lower Kent Ridge Road, 119074, Singapore

Tóm tắt

Abstract Background

Exosomes or secreted bi-lipid vesicles from human ESC-derived mesenchymal stem cells (hESC-MSCs) have been shown to reduce myocardial ischemia/reperfusion injury in animal models. However, as hESC-MSCs are not infinitely expansible, large scale production of these exosomes would require replenishment of hESC-MSC through derivation from hESCs and incur recurring costs for testing and validation of each new batch. Our aim was therefore to investigate if MYC immortalization of hESC-MSC would circumvent this constraint without compromising the production of therapeutically efficacious exosomes.

 Methods

The hESC-MSCs were transfected by lentivirus carrying a MYC gene. The transformed cells were analyzed for MYC transgene integration, transcript and protein levels, and surface markers, rate of cell cycling, telomerase activity, karyotype, genome-wide gene expression and differentiation potential. The exosomes were isolated by HPLC fractionation and tested in a mouse model of myocardial ischemia/reperfusion injury, and infarct sizes were further assessed by using Evans' blue dye injection and TTC staining.

 Results

MYC-transformed MSCs largely resembled the parental hESC-MSCs with major differences being reduced plastic adherence, faster growth, failure to senesce, increased MYC protein expression, and loss of in vitro adipogenic potential that technically rendered the transformed cells as non-MSCs. Unexpectedly, exosomes from MYC-transformed MSCs were able to reduce relative infarct size in a mouse model of myocardial ischemia/reperfusion injury indicating that the capacity for producing therapeutic exosomes was preserved.

 Conclusion

Our results demonstrated that MYC transformation is a practical strategy in ensuring an infinite supply of cells for the production of exosomes in the milligram range as either therapeutic agents or delivery vehicles. In addition, the increased proliferative rate by MYC transformation reduces the time for cell production and thereby reduces production costs.

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Journal of Translational Medicine

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