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Pre-Differentiation of KIND1 Human Embryonic Stem Cell Line into Type I Alveolar Epithelial Cells Alleviates Symptoms of Idiopathic Pulmonary Fibrosis in Mice

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Abstract

Stem cell therapy uses of the ability of stem cells to differentiate into cells of other lineages, engraft into damaged tissues and promote their regeneration. However, the use of ESCs has been limited due to safety issues, which we have attempted to overcome by differentiating the cells before transplantation. In this study, we have induced the differentiation of KIND1 hESC line using a cocktail of growth factors present in SAGM. These differentiated cells were then transplanted into a bleomycin-induced mouse model of IPF, a degenerative lung disease with no permanent cure, to determine whether they could reduce the symptoms of the disease. We found that KIND1 hESCs, grown in SAGM for 48 h, predominantly differentiated into AQP5+ type-I alveolar epithelial cells, and a small fraction of SP-C+ AE-II cells. When transplanted into mice with IPF, these differentiated cells showed a significant alleviation of the disease symptoms, by engrafting into, and promoting regeneration of, the damaged tissue, thereby restoring the gas exchange capacity of the lungs. Thus, pre-differentiating hESCs into a specific lineage before transplantation could provide a safer and more economical therapy for degenerative diseases like IPF.

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Abbreviations

SC:

Stem cell

MSC:

Mesenchymal stem cells

ESC:

Embryonic stem cell

CD:

Cluster of differentiation

ECM:

Extracellular matrix

bFGF:

Basic fibroblast growth factor

AE cells:

Alveolar epithelial cells

AE-I:

Type I alveolar epithelial cell

AE-II:

Type II alveolar epithelial cell

AQP:

Aquaporin

SP-C:

Surfactant protein-C

SAGM:

Small airway epithelial cell growth medium

IPF:

Idiopathic pulmonary fibrosis

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

Bleo:

Bleomycin

PB:

Peripheral blood

BALF:

Bronchoalveolar lavage fluid

SEM:

Standard error of mean

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Acknowledgements

We are greatly obliged to Dr. Sujata Mohanty (Professor, Centre of Excellence for Stem Cell Research, All India Institute of Medical Sciences, New Delhi, India) and her research fellows for their endless help in establishing the cell line and in allowing us to perform some of the experiments in their laboratory. We also acknowledge the Indian Institute of Chemical Biology (IICB), Kolkata, for allowing us to use the BD LSRFortessa. We would like to acknowledge the Indian Council of Medical Research (ICMR), UGC-UPE-II and UGC-DRS-SAP-Phase II for providing research support for this work, DST-FIST-Phase III for departmental instruments, and DST-PURSE-Phase II for departmental infrastructural support. We acknowledge the University Grants Commission (UGC), New Delhi, for providing fellowship to SM.

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  1. Immunology and Regenerative Medicine Research Laboratory, Department of Zoology, University of Calcutta, 35, Ballygunge Circular Road, Kolkata, West Bengal, 700019, India

    Shinjini Mitra & Ena Ray Banerjee

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  1. Shinjini Mitra
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All authors contributed to the study conception and design. Conceptualization and supervision was done by ERB. Study design, material preparation, data collection and analysis were performed by SM. The first draft of the manuscript was written by SM and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Ena Ray Banerjee.

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Mitra, S., Banerjee, E.R. Pre-Differentiation of KIND1 Human Embryonic Stem Cell Line into Type I Alveolar Epithelial Cells Alleviates Symptoms of Idiopathic Pulmonary Fibrosis in Mice. Proc Zool Soc 76, 73–90 (2023). https://doi.org/10.1007/s12595-023-00469-2

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