1H-NMR spectroscopy shows cellular uptake of HEPES buffer by human cell lines—an effect to be considered in cell culture experiments

Depping, Reinhard; Seeger, Karsten

doi:10.1007/s00216-018-1518-4

Cited by 8 publications

(10 citation statements)

References 14 publications

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“…Nevertheless, emerging severe adverse effects in cell/tissue cultures have been reported. For example, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) was found to be endocytosed by cells [ 8 ]. RPMI 1640 medium supplied with HEPES (25 mM) resulted in significant cytotoxicity under visible light [ 9 ], while the presence of HEPES in the culture medium clearly up-regulate lysosomal-autophagic aggravation and inflammatory signaling of the cells [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Zwitterionic betaines over HEPES as the new generation biocompatible pH buffers for cell culture

Liu

Sun

Wan

et al. 2023

Bioactive Materials

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Section: Introductionmentioning

confidence: 99%

Zwitterionic betaines over HEPES as the new generation biocompatible pH buffers for cell culture

Liu

Sun

Wan

et al. 2023

Bioactive Materials

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“…Increase buffering capacity of medium to maintain the pH of cultures [50] Phenol Red pH indicator [29] 2-2-1 FBS containing medium formulation FBS-containing medium consisted of, 10% FBS (Bovogen), 1% Anti-Anti. To induce osteogenic differentiation of the cells, this medium was supplemented with 50 μg/mL ascorbic-acid-2-phosphate (Sigma-Aldrich, A8960), 100 nM dexamethasone (Sigma-Aldrich, D4902), 10 mM β-glycerophosphate 2-4 Cell isolation, expansion, and subsequent seeding hBMSCs were isolated from 2 donors of human bone marrow (Lonza, USA) and characterized as previously described [52].…”

Section: Hepesmentioning

confidence: 99%

“…Basic building blocks of proteins and essential for cell proliferation and viability [45] Vitamins Essential for cell growth and proliferation, act as co-factor for enzymes [7,46] Inorganic salts Maintain the osmotic balance and regulate membrane potential which is necessary to transport ions through cell membrane [47,48] Glucose Main source of energy for cell metabolism [29] Sodium Pyruvate By-product of glucose consumption in the glycolytic pathway and functions as a source of energy [49] HEPES Increase buffering capacity of medium to maintain the pH of cultures [50] Phenol Red pH indicator [29] 2-2-1 FBS containing medium formulation FBS-containing medium consisted of, 10% FBS (Bovogen), 1% Anti-Anti. To induce osteogenic differentiation of the cells, this medium was supplemented with 50 μg/mL ascorbic-acid-2-phosphate (Sigma-Aldrich, A8960), 100 nM dexamethasone (Sigma-Aldrich, D4902), 10 mM β-glycerophosphate 2-4 Cell isolation, expansion, and subsequent seeding hBMSCs were isolated from 2 donors of human bone marrow (Lonza, USA) and characterized as previously described [52].…”

Section: Amino Acidsmentioning

confidence: 99%

Development of serum substitute medium for bone tissue engineering

Ansari

Ito

Hofmann

2022

Preprint

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In tissue engineering, cells are grown often on scaffolds and subjected to chemical/mechanical stimuli. Most such cultures still use fetal bovine serum (FBS) despite its known disadvantages including ethical concerns, safety issues, and variability in composition, which greatly influences the experimental outcomes. To overcome the disadvantages of using FBS, chemically defined serum substitute medium needs to be developed. Development of such medium depends on cell type and application - which makes it impossible to define one universal serum substitute medium for all cells in any application. Here, we developed a serum substitute medium for bone tissue engineering (BTE) in a step-by-step process. Essential components were added to the medium while human bone marrow mesenchymal stromal cells (hBMSCs, osteoblast progenitor cells) were cultured in two-dimensional (2D) and three-dimensional (3D) substrates. In a 3-week culture, the developed serum substitute medium worked equally well as FBS containing medium in term of cell attachment to the substrate, cell survival, osteoblast differentiation, and deposition of extracellular matrix. In the next step, the use of serum substitute medium was evaluated when culturing cells under mechanical loading in the form of shear stress. The outcomes showed that the application of shear stress is essential to improve extracellular matrix formation while using serum substitute medium. The developed serum substitute medium could pave the way in replacing FBS for BTE studies eliminating the use of controversial FBS and providing a better-defined chemical environment for BTE studies.

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“…Briefly, HEPES activated lysosomal transcription factors in macrophages ( Tol et al, 2018 ), inhibited the prion protein conversion in neural stem cells and affected their viability and differentiation ( Delmouly et al, 2011 ). Another study showed cellular uptake of HEPES in human cell lines (MCF-7, U2OS, HeLa) that persisted for 48 h after cells were returned to HEPES-free media ( Depping and Seeger, 2019 ).…”

Section: The Problemsmentioning

confidence: 99%

Toward Best Practices for Controlling Mammalian Cell Culture Environments

Klein

Steckbauer

Alsolami

et al. 2022

Front. Cell Dev. Biol.

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The characterization, control, and reporting of environmental conditions in mammalian cell cultures is fundamental to ensure physiological relevance and reproducibility in basic and preclinical biomedical research. The potential issue of environment instability in routine cell cultures in affecting biomedical experiments was identified many decades ago. Despite existing evidence showing variable environmental conditions can affect a suite of cellular responses and key experimental readouts, the underreporting of critical parameters affecting cell culture environments in published experiments remains a serious problem. Here, we outline the main sources of potential problems, improved guidelines for reporting, and deliver recommendations to facilitate improved culture-system based research. Addressing the lack of attention paid to culture environments is critical to improve the reproducibility and translation of preclinical research, but constitutes only an initial step towards enhancing the relevance of in vitro cell cultures towards in vivo physiology.

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1H-NMR spectroscopy shows cellular uptake of HEPES buffer by human cell lines—an effect to be considered in cell culture experiments

Cited by 8 publications

References 14 publications

Zwitterionic betaines over HEPES as the new generation biocompatible pH buffers for cell culture

Zwitterionic betaines over HEPES as the new generation biocompatible pH buffers for cell culture

Development of serum substitute medium for bone tissue engineering

Toward Best Practices for Controlling Mammalian Cell Culture Environments

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