Webinar: Controlling HIF-1α: The Secret for Optimal MSC Growth

Abstract:

Mesenchymal Stromal/Stem Cells (MSCs) are in the news as clinical trials open up for MSC-based COVID treatments, thus maintaining optimal MSC growth conditions in vitro has never been more important. Human bone marrow MSCs originate in vivobetween 1-5% oxygen (O2).¹ When human MSC (hMSC) are used experimentally, they are incubated in physioxic conditions in vitro (3% O₂) but are often handled at supraphysioxic levels in room air (20% O2) in biological safety cabinets (BSC) for media changes or culture treatments. BioSpherix previously reported that cells kept under constant 3% O₂, for both culture and handling, resulted in an increased hMSC yield over those with brief exposure to room air.²

Hypoxia-Inducible Factor 1 alpha (HIF-1α) is upregulated in low oxygen environments and acts as a transcription factor by binding to the hypoxia response element region of target genes. Conversely, HIF-1α is degraded when exposed to oxygen. Downstream of HIF-1α are genes involved in cell growth and proliferation, important for hMSC differentiation and function. 

Here, we examined protein concentration changes in hMSCs after exposure to room air conditions to assess how cell handling environments regulate HIF-1α expression in hMSCs. We hypothesized that even brief exposure to room air during routine cell handling would reduce HIF-1α expression in hMSCs when compared to cells maintained in physioxic conditions and would regulate downstream targets accordingly.

Learning Objectives:

• Understand the molecular changes in MSCs resulting from fluctuating oxygen conditions.
• Find out how the automated Western blotting platforms Wes™ and Jess™ are used to reproducibly quantify protein abundance in MSCs after a brief exposure to room air.
• Discover the latest technologies for improving MSC growth in culture by controlling HIF-1α protein levels.