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Quality Media for Successful Cell Cultivation

February 4, 2021 / by Lifeline® Cell Technology

The Cell Culture Media Makes the Culture Environment

In order to successfully cultivate cells in the lab, researchers must use a suitable culture environment with the necessary nutrients for survival and proliferation, which is largely supplied by the cell culture medium. It also helps to regulate physiochemical parameters, such as pH, osmotic pressure and the O2/CO2 tension of the cell culture environment. Using suboptimal culture media can produce varying results from the expression of aberrant phenotypes to a complete failure of the cell culture. At Lifeline®, we offer cell culture media optimized for growth of many different cell types isolated from primary tissues as well as media kits for the maintenance or differentiation of undifferentiated stem cells. Our media does not contain antimicrobials or the pH indicator phenol red, known to weakly activate the estrogen receptor, which can interfere with experiments investigating hormone signaling pathways.

Several recent publications demonstrate the utility of Lifeline culture media for the isolation and expansion of human tracheobronchial epithelial cells (BronchiaLife Complete Medium) and foreskin-derived dermal microvascular endothelial cells (VascuLife EnGS Complete Medium) for their research, which we have summarized here.

Studies using Lifeline Medium to Culture Human Primary Cells

The first study by Dakhama and Colleagues used Lifeline’s BronchiaLife Complete Medium to isolate and grow normal human tracheobronchial epithelial cells (HBTE’s) to determine the mechanism by which Toll-interacting protein (Tollip) regulates the proinflammatory cytokine responses (i.e., IL-13 and IL-33) induced by rhinovirus (RV) infection. IL-8, IRAK1 and soluble ST2 and their interactions with Tollip play key roles in this response but the exact mechanisms were not clearly understood. The authors utilized knockdown and knockout HTBE cells generated by shRNA knockdown and CRISPR/Cas9 approaches, respectively, to assess IL-8 levels after stimulation with type 2 proinflammatory cytokines IL-13 and IL-33 during RV infection.

The authors found that Tollip-deficient HTBE cells had reduced levels of secreted ST2 but produced excessive amounts of IL-8 compared to Tollip-sufficient cells, which is directly linked to IRAK1 activation. These findings led the researchers to propose a novel mechanism where, in the presence of IL-13 and IL-33, Tollip promotes soluble ST2 production and inhibits IRAK1 activation, which together restrict excess IL-8 production and the development of detrimental inflammation. This suggests a combinative therapeutic approach – blocking IRAK1 and adding sST2 – may be potential avenues to prevent excessive airway inflammation during RV-mediated asthma exacerbations.

The second study published by Groeber and Colleagues details the establishment of a novel protocol to generate microvascularized skin equivalents for clinical and research applications. The lack of functional vasculature in the engineered skin grafts currently used requires the formation of new blood vessels through angiogenesis, which impairs graft integration and increases the potential for graft rejection. As well, In vitro non-vascularized skin models are of limited value with regard to their ability to reflect the physiological conditions of a full organ.

Here, the researchers used a biological vascularized scaffold (BioVaSc) in combination with a tailor-made bioreactor system to generate skin equivalents with a perfused vascular network. To form the vascular network, human dermal microvascular endothelial cells (hDMEC) from human foreskin tissue (isolated and cultured in Lifeline VascuLife EnGS Complete medium) were seeded into the acellular vascular structures of the BioVaSc. The surface of the BioVaSc matrix was inoculated with human epidermal keratinocytes (hEK), human dermal fibroblasts (hDF) to establish a well-stratified epidermal layer at the air-liquid-interface of the specific-designed bioreactor. Hematoxylin & eosin and immunohistological staining confirmed the specific histological architecture representative of the human skin layers. As well, the endothelial cells lining the vessels in BioVaSc could be perfused with a physiologically relevant fluidic flow. Taken together, this fully vascularized, functional human skin model represents the in vivo conditions of a full organ and has the potential for use in skin grafting to improve clinical outcomes and further dermatological research by representing a more biomimetic in vitro model.

Are you using Lifeline cells and/or culture media in your research? Your publication could be featured here on the blog!

For more information on these and Lifeline’s other culture media products, please visit our website:

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