On-line First

J Plant Biotechnol

Published online July 5, 2024

© The Korean Society of Plant Biotechnology

Production of human epidermal growth factor (hEGF) in cell suspension cultures of Centella asiatica (L.) Urban using balloon-type bubble bioreactors

Ga-Eun Baek 1, Han-Sol Lee 1, Xinlei Bai 1, Hosakatte Niranjana Murthy 1, 2, Eun-Jeong Son 3, Hyo Hyun Seo 4, Sang Hyum Moh 4, So-Young Park 1*

1Chungbuk National University, 2Karnatak University, 3Korea Institute of Oriental Medicine, 4Plant Cell Research Institute of BIO-FD&C Co., Ltd.

Received: 14 June 2024; Revised: 3 July 2024; Accepted: 5 July 2024

Abstract

The human epidermal growth factor (hEGF) has several medical and pharmacological uses. The goal of this study was to determine whether the medicinal plant Centella asiatica callus and cell suspension cultures could produce hEGF. The callus of C. asiatica was transformed using Agrobacterium tumefaciens strains carrying the hEGF gene and the hEGF-KDEL gene (KDEL sequence targeting peptides to endoplasmic reticulum) on plasmids pKRE1 and pKRE2, respectively, under the control of 35S promoters. Polymerase chain reaction with gene-specific primers was used to select transformed callus lines. The qRT-PCR and Western blot methods were used to confirm that the EGF gene was expressed in transformed lines. In balloon-type bubble bioreactors, cell suspension cultures were established using transformed line EK2 cells. In bioreactor cultures, efficient transformed cell biomass could be produced. The hEGF protein isolated from transformed cells has also demonstrated in vitro cell proliferation of human keratinocytes (HaCaT) cells. These findings showed that plant expression systems, particularly plant cell cultures, could produce recombinant hEGF.

Keywords Bioreactor cultures, Centella asiatica, cell suspension culture, human epidermal growth factor, recombinant protein

Article

On-line First

J Plant Biotechnol

Published online July 5, 2024

Copyright © The Korean Society of Plant Biotechnology.

Production of human epidermal growth factor (hEGF) in cell suspension cultures of Centella asiatica (L.) Urban using balloon-type bubble bioreactors

Ga-Eun Baek 1, Han-Sol Lee 1, Xinlei Bai 1, Hosakatte Niranjana Murthy 1, 2, Eun-Jeong Son 3, Hyo Hyun Seo 4, Sang Hyum Moh 4, So-Young Park 1*

1Chungbuk National University, 2Karnatak University, 3Korea Institute of Oriental Medicine, 4Plant Cell Research Institute of BIO-FD&C Co., Ltd.

Received: 14 June 2024; Revised: 3 July 2024; Accepted: 5 July 2024

Abstract

The human epidermal growth factor (hEGF) has several medical and pharmacological uses. The goal of this study was to determine whether the medicinal plant Centella asiatica callus and cell suspension cultures could produce hEGF. The callus of C. asiatica was transformed using Agrobacterium tumefaciens strains carrying the hEGF gene and the hEGF-KDEL gene (KDEL sequence targeting peptides to endoplasmic reticulum) on plasmids pKRE1 and pKRE2, respectively, under the control of 35S promoters. Polymerase chain reaction with gene-specific primers was used to select transformed callus lines. The qRT-PCR and Western blot methods were used to confirm that the EGF gene was expressed in transformed lines. In balloon-type bubble bioreactors, cell suspension cultures were established using transformed line EK2 cells. In bioreactor cultures, efficient transformed cell biomass could be produced. The hEGF protein isolated from transformed cells has also demonstrated in vitro cell proliferation of human keratinocytes (HaCaT) cells. These findings showed that plant expression systems, particularly plant cell cultures, could produce recombinant hEGF.

Keywords: Bioreactor cultures, Centella asiatica, cell suspension culture, human epidermal growth factor, recombinant protein

JPB
Vol 51. 2024

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Plant Biotechnology

pISSN 1229-2818
eISSN 2384-1397
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