Baker M. 1,500 scientists lift the lid on reproducibility. Nature. 2016;533(7604):452–4.

Article  PubMed  CAS  Google Scholar 

Beachy SH, Nair L, Laurencin C, Tsokas KA, Lundberg MS. Sources of variability in clinical translation of regenerative engineering products: insights from the National Academies Forum on Regenerative Medicine. Regenerative Engineering and Translational Medicine. 2020;6(1):1–6.

Article  CAS  Google Scholar 

Klein SG, Alsolami SM, Steckbauer A, Arossa S, Parry AJ, Ramos Mandujano G, et al. A prevalent neglect of environmental control in mammalian cell culture calls for best practices. Nat Biomed Eng. 2021;5(8):787–92.

Article  PubMed  Google Scholar 

•• Klein SG, Steckbauer A, Alsolami SM, Arossa S, Parry AJ, Li M, et al. Toward best practices for controlling mammalian cell culture environments. Frontiers in Cell and Developmental Biology. 2022;10. This article calls for new processes for reporting cell culture conditions for reproducibility and offers specific guidelines.

Mantel CR, O’Leary HA, Chitteti BR, Huang X, Cooper S, Hangoc G, et al. Enhancing Hematopoietic stem cell transplantation efficacy by mitigating oxygen shock. Cell. 2015;161(7):1553–65.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Broxmeyer HE, O’Leary HA, Huang X, Mantel C. The importance of hypoxia and extra physiologic oxygen shock/stress for collection and processing of stem and progenitor cells to understand true physiology/pathology of these cells ex vivo. Curr Opin Hematol. 2015;22(4):273–8.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Ast T, Mootha VK. Oxygen and mammalian cell culture: are we repeating the experiment of Dr. Ox? Nature metabolism. 2019;1(9):858–60.

DiProspero TJ, Dalrymple E, Lockett MR. Physiologically relevant oxygen tensions differentially regulate hepatotoxic responses in HepG2 cells. Toxicology in vitro: an international journal published in association with BIBRA. 2021;105156.

Kumar A, Dailey LA, Swedrowska M, Siow R, Mann GE, Vizcay-Barrena G, et al. Quantifying the magnitude of the oxygen artefact inherent in culturing airway cells under atmospheric oxygen versus physiological levels. FEBS Lett. 2016;590(2):258–69.

Article  PubMed  CAS  Google Scholar 

Kumar B, Adebayo AK, Prasad M, Capitano ML, Wang R, Bhat-Nakshatri P, et al. Tumor collection/processing under physioxia uncovers highly relevant signaling networks and drug sensitivity. Sci Adv. 2022;8(2):eabh3375.

• Mount S, Kanda P, Parent S, Khan S, Michie C, Davila L, et al. Physiologic expansion of human heart-derived cells enhances therapeutic repair of injured myocardium. Stem Cell Res Ther. 2019;10(1):1–16. This article shows that protecting cells from uncontrolled room air during cGMP processing enhances engraftment in vivo.

Anderson DE, Markway BD, Weekes KJ, McCarthy HE, Johnstone B. Physioxia promotes the articular chondrocyte-like phenotype in human chondroprogenitor-derived self-organized tissue. Tissue Eng Part A. 2018;24(3–4):264–74.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Shimoni YFT, Srinivasan V, Szeto R. Reducing variability in cell-specific productivity in perfusion culture: a case study. Bioprocess International. 2018.

Shimoni Y, Pathange LP. Product quality attribute shifts in perfusion systems, part 2: elucidating cellular mechanisms. BioProcess International. 2020.

Gurvich OL, Puttonen KA, Bailey A, Kailaanmaki A, Skirdenko V, Sivonen M, et al. Transcriptomics uncovers substantial variability associated with alterations in manufacturing processes of macrophage cell therapy products. Sci Rep. 2020;10(1):14049.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Burke CJ, Zylberberg C. Sources of Variability in Manufacturing of Cell Therapeutics. Regenerative engineering and translational medicine. 2019;5(4):332–40.

Article  Google Scholar 

Stover AE, Herculian S, Banuelos MG, Navarro SL, Jenkins MP, Schwartz PH. Culturing human pluripotent and neural stem cells in an enclosed cell culture system for basic and preclinical research. J Vis Exp. 2016(112).

Drexler HG, Dirks WG, MacLeod RA, Uphoff CC. False and mycoplasma-contaminated leukemia-lymphoma cell lines: time for a reappraisal. Int J Cancer. 2017;140(5):1209–14.

Article  PubMed  CAS  Google Scholar 

Chang Y, Goldberg VM, Caplan AI. Toxic effects of gentamicin on marrow-derived human mesenchymal stem cells. Clin Orthop Relat Res. 2006;452:242–9.

Article  PubMed  Google Scholar 

Cohen S, Samadikuchaksaraei A, Polak JM, Bishop AE. Antibiotics reduce the growth rate and differentiation of embryonic stem cell cultures. Tissue Eng. 2006;12(7):2025–30.

Article  PubMed  CAS  Google Scholar 

Jiang X, Baucom C, Elliott RL. Mitochondrial toxicity of azithromycin results in aerobic glycolysis and DNA damage of human mammary epithelia and fibroblasts. Antibiotics (Basel, Switzerland). 2019;8(3).

Relier S, Yazdani L, Ayad O, Choquet A, Bourgaux JF, Prudhomme M, et al. Antibiotics inhibit sphere-forming ability in suspension culture. Cancer Cell Int. 2016;16:6.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Ryu AH, Eckalbar WL, Kreimer A, Yosef N, Ahituv N. Use antibiotics in cell culture with caution: genome-wide identification of antibiotic-induced changes in gene expression and regulation. Sci Rep. 2017;7(1):7533.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Varghese DS, Parween S, Ardah MT, Emerald BS, Ansari SA. Effects of aminoglycoside antibiotics on human embryonic stem cell viability during differentiation in vitro. Stem Cells Int. 2017;2017:2451927.

Article  PubMed  PubMed Central  CAS  Google Scholar 

He Y, Darou S, Henn S, Walter R, Cundell A, Yerden R, et al. Temperature and relative humidity control to reduce bioburden in a closed cell processing and production system without disinfectants. BioPharm International. 2021.

Carrel A, Lindbergh CA. The culture of organs. Am J Med Sci. 1938;196(5):732.

Article  Google Scholar 

Kleman AM, Yuan JY, Aja S, Ronnett GV, Landree LE. Physiological glucose is critical for optimized neuronal viability and AMPK responsiveness in vitro. J Neurosci Methods. 2008;167(2):292–301.

Article  PubMed  CAS  Google Scholar 

Davidson MD, Pickrell J, Khetani SR. Physiologically inspired culture medium prolongs the lifetime and insulin sensitivity of human hepatocytes in micropatterned co-cultures. Toxicology. 2021;449: 152662.

Article  PubMed  CAS  Google Scholar 

Moradi F, Moffatt C, Stuart JA. The effect of oxygen and micronutrient composition of cell growth media on cancer cell bioenergetics and mitochondrial networks. Biomolecules. 2021;11(8):1177.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Rogers LK, Cismowski MJ. Oxidative stress in the lung - the essential paradox. Current opinion in toxicology. 2018;7:37–43.

Article  PubMed  Google Scholar 

Keeley TP, Mann GE. Defining physiological normoxia for improved translation of cell physiology to animal models and humans. Physiol Rev. 2019;99(1):161–234.

Article  PubMed  CAS  Google Scholar 

Carreau A, El Hafny-Rahbi B, Matejuk A, Grillon C, Kieda C. Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia. J Cell Mol Med. 2011;15(6):1239–53.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Wenger RH, Kurtcuoglu V, Scholz CC, Marti HH, Hoogewijs D. Frequently asked questions in hypoxia research. Hypoxia. 2015;3:35–43.

Article  PubMed  PubMed Central  Google Scholar 

Bambrick L, Kostov Y, Rao G. In vitro cell culture pO2 is significantly different from incubator pO2. Biotechnol Prog. 2011;27(4):1185–9.

Article  PubMed  CAS  Google Scholar 

Mohyeldin A, Garzon-Muvdi T, Quinones-Hinojosa A. Oxygen in stem cell biology: a critical component of the stem cell niche. Cell Stem Cell. 2010;7(2):150–61.

Article  PubMed  CAS  Google Scholar 

Shariati L, Esmaeili Y, Javanmard SH, Bidram E, Amini A. Organoid technology: current standing and future perspectives. Stem Cells. 2021.

Bannier-Helaouet M, Post Y, Korving J, Trani Bustos M, Gehart H, Begthel H, et al. Exploring the human lacrimal gland using organoids and single-cell sequencing. Cell Stem Cell. 2021.

Li Q, Qi G, Liu X, Bai J, Zhao J, Tang G, et al. Universal peptide hydrogel for scalable physiological formation and bioprinting of 3D spheroids from human induced pluripotent stem cells. Adv Funct Mater. 2021;31(41):2104046.

Lehmann R, Lee CM, Shugart EC, Benedetti M, Charo RA, Gartner Z, et al. Human organoids: a new dimension in cell biology. Mol Biol Cell. 2019;30(10):1129–37.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Kim J, Koo B-K, Knoblich JA. Human organoids: model systems for human biology and medicine. Nat Rev Mol Cell Biol. 2020;21(10):571–84.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Thippabhotla S, Zhong C, He M. 3D cell culture stimulates the secretion of in vivo like extracellular vesicles. Sci Rep. 2019;9(1):1–14.

Article  CAS  Google Scholar 

Fioretta ES, Simonet M, Smits AI, Baaijens FP, Bouten CV. Differential response of endothelial and endothelial colony forming cells on electrospun scaffolds with distinct microfiber diameters. Biomacromol. 2014;15(3):821–9.

Article  CAS  Google Scholar 

Wright Muelas M, Ortega F, Breitling R, Bendtsen C, Westerhoff HV. Rational cell culture optimization enhances experimental reproducibility in cancer cells. Sci Rep. 2018;8(1):3029.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Tse HM, Gardner G, Dominguez-Bendala J, Fraker CA. The importance of proper oxygenation in 3D culture. Front Bioeng Biotechnol. 2021;9(241): 634403.

Article  PubMed  PubMed Central  Google Scholar 

Wohlrab P, Johann Danhofer M, Schaubmayr W, Tiboldi A, Krenn K, Markstaller K, et al. Oxygen conditions oscillating between hypoxia and hyperoxia induce different effects in the pulmonary endothelium compared to constant oxygen conditions. Physiol Rep. 2021;9(3): e14590.

Article  PubMed  PubMed Central  CAS  Google Scholar 

Shin DY, Huang X, Gil CH, Aljoufi A, Ropa J, Broxmeyer HE. Physioxia enhances T-cell development ex vivo from human hematopoietic stem and progenitor cells. Stem Cells. 2020

Sagan DS, Carl Margulis L. Life. Encyclopedia Britannica 2020

Hofer M, Lutolf MP. Engineering organoids. Nat Rev Mater. 2021;1–19.

Bhatia SN, Ingber DE. Microfluidic organs-on-chips. Nat Biotechnol. 2014;32(8):760–72.

Article