Human Embryonic Kidney 293 Cells: A Vehicle for Biopharmaceutical Manufacturing, Structural Biology, and Electrophysiology

Hu, Jiayin; Han, Jizhong; Li, Haoran; Zhang, Xian; Liu, Lan Lan; Chen, Fei; Zeng, Bin

doi:10.1159/000485501

Cited by 75 publications

(64 citation statements)

References 69 publications

(86 reference statements)

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“…These immortalized cells come from Human Embryonic Kidney (34) and are used in many laboratories for heterologous expression of proteins. Previous heterologous expression systems were not of human origin and, accordingly, the development of HEK-293T cells was a of paramount relevance for biomolecular research; they are currently used in biochemistry, pharmacology, electrophysiology and biotechnology approaches aimed at advancing knowledge into protein structure/function relationships (35)(36)(37). Cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 2 mM L-glutamine, 100 U/ml penicillin/streptomycin, and 5% (v/v) heat inactivated fetal bovine serum (FBS) (Invitrogen, Paisley, Scotland, United Kingdom).…”

Section: Cell Culture and Transient Transfectionmentioning

confidence: 99%

Specificity and nanomolar potency of melatonin on G-protein coupled melatonin MT1 and MT2 receptors expressed in HEK-293T human embryo kidney cells

Rivas‐Santisteban¹,

Reyes‐Resina²,

Raïch³

et al. 2019

Melatonin Res.

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This is a pre-registered study, i.e. a study whose hypotheses and experiments designed to address these hypotheses have been deposited in a database before starting the experiments. The study aims at assessing the Gs versus Gi coupling and the potency of melatonin in the human version of melatonin MT1 and MT2 G-protein-coupled receptors expressed in HEK-293T cells. The results show that these receptors are Gi but not Gs coupled. By using a standard procedure of modulation of 0.5 µM forskolin-induced cAMP levels, it was found that the potency on MT2 receptor-mediated actions is in the low nanomolar range, but the potency on MT1 receptor is in the high nanomolar range. The potency of melatonin to stimulate the MT2 receptor is similar to that of a selective agonist, N-[2-(2-methoxy-6H-isoindolo[2,1-a]indol-11-yl)ethyl]butanamide (IIK7). Overall, the data on the potency of melatonin on its receptors will provide a new look for melatonin research. It is important to consider this finding for appropriately addressing physiological or therapeutic effects based on melatonin potency. Thus, the low doses of melatonin used in the existing prolonged release preparations or in other supplements should be revisited.

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Section: Cell Culture and Transient Transfectionmentioning

confidence: 99%

Specificity and nanomolar potency of melatonin on G-protein coupled melatonin MT1 and MT2 receptors expressed in HEK-293T human embryo kidney cells

Rivas‐Santisteban¹,

Reyes‐Resina²,

Raïch³

et al. 2019

Melatonin Res.

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“…PTM is present in mammalian cell lines; however, their glycosylation pattern is different from human-type glycosylation. HEK293, HKB11, PER.C6, HeLa, and CAP cells (all human cells lines) are being also studied for protein expression (Bandaranayake and Almo, 2014;Dumont et al, 2016;Dyson, 2016;Hu et al, 2018;Gupta S. K. et al, 2019;Hunter et al, 2019). A human cell line enhances the expression of proteins with human-like PTMs.…”

Section: Mammalian Cellsmentioning

confidence: 99%

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Tripathi

Shrivastava

2019

Front. Bioeng. Biotechnol.

400

243

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Infectious diseases, along with cancers, are among the main causes of death among humans worldwide. The production of therapeutic proteins for treating diseases at large scale for millions of individuals is one of the essential needs of mankind. Recent progress in the area of recombinant DNA technologies has paved the way to producing recombinant proteins that can be used as therapeutics, vaccines, and diagnostic reagents. Recombinant proteins for these applications are mainly produced using prokaryotic and eukaryotic expression host systems such as mammalian cells, bacteria, yeast, insect cells, and transgenic plants at laboratory scale as well as in large-scale settings. The development of efficient bioprocessing strategies is crucial for industrial production of recombinant proteins of therapeutic and prophylactic importance. Recently, advances have been made in the various areas of bioprocessing and are being utilized to develop effective processes for producing recombinant proteins. These include the use of high-throughput devices for effective bioprocess optimization and of disposable systems, continuous upstream processing, continuous chromatography, integrated continuous bioprocessing, Quality by Design, and process analytical technologies to achieve quality product with higher yield. This review summarizes recent developments in the bioprocessing of recombinant proteins, including in various expression systems, bioprocess development, and the upstream and downstream processing of recombinant proteins.

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“…As for cardiac genetic diseases is concerned, pathological mutations have often been found in genes encoding junctional proteins, cytoskeletal components, signaling molecules, ion channels and accessory proteins. In most cases, heterologous systems allowed the heterologous expression of those proteins, either Wild Type (WT) or mutant, for a functional evaluation of the genetic alterations [5].…”

Section: Non-cardiovascular Cellsmentioning

confidence: 99%

Human Cell Modeling for Cardiovascular Diseases

Lippi

Stadiotti

Pompilio

et al. 2020

IJMS

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The availability of appropriate and reliable in vitro cell models recapitulating human cardiovascular diseases has been the aim of numerous researchers, in order to retrace pathologic phenotypes, elucidate molecular mechanisms, and discover therapies using simple and reproducible techniques. In the past years, several human cell types have been utilized for these goals, including heterologous systems, cardiovascular and non-cardiovascular primary cells, and embryonic stem cells. The introduction of induced pluripotent stem cells and their differentiation potential brought new prospects for large-scale cardiovascular experiments, bypassing ethical concerns of embryonic stem cells and providing an advanced tool for disease modeling, diagnosis, and therapy. Each model has its advantages and disadvantages in terms of accessibility, maintenance, throughput, physiological relevance, recapitulation of the disease. A higher level of complexity in diseases modeling has been achieved with multicellular co-cultures. Furthermore, the important progresses reached by bioengineering during the last years, together with the opportunities given by pluripotent stem cells, have allowed the generation of increasingly advanced in vitro three-dimensional tissue-like constructs mimicking in vivo physiology. This review provides an overview of the main cell models used in cardiovascular research, highlighting the pros and cons of each, and describing examples of practical applications in disease modeling.

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Human Embryonic Kidney 293 Cells: A Vehicle for Biopharmaceutical Manufacturing, Structural Biology, and Electrophysiology

Cited by 75 publications

References 69 publications

Specificity and nanomolar potency of melatonin on G-protein coupled melatonin MT1 and MT2 receptors expressed in HEK-293T human embryo kidney cells

Specificity and nanomolar potency of melatonin on G-protein coupled melatonin MT1 and MT2 receptors expressed in HEK-293T human embryo kidney cells

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Human Cell Modeling for Cardiovascular Diseases

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