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The gold standards for coverage of wounds that cannot be primarily closed are full thickness skin grafts (FTSGs) and split thickness skins graft (STSGs). FTSGs harvest sites generally require primary closure, which limits availability, especially when treating larger wounds. STSGs have many shortcomings, including donor site morbidity. Fractional autologous skin replacement can be utilized in conjunction with or in lieu of STSGs to both improve graft outcomes of large wounds and to decrease donor site morbidity. Skin can be mechanically or chemically fractionated. Fractionated skin can be advantageous, as adnexal structures provide additional functionality without donor site morbidity. In this review, we will discuss current and emerging techniques in fractional skin replacement. Full article

The assessment of human liver stem cells (HLSCs) as cell therapeutics requires scalable, controlled expansion processes. We first focused on defining appropriate process parameters for HLSC expansion such as seeding density, use of antibiotics, optimal cell age and critical metabolite concentrations in conventional 2D culture systems. For scale-up, we transferred HLSC expansion to multi-plate and stirred-tank bioreactor systems to determine their limitations. A seeding density of 4000 cells cm⁻² was needed for efficient expansion. Although growth was not significantly affected by antibiotics, the concentrations of lactate and ammonia were important. A maximum expansion capacity of at least 20 cumulative population doublings (cPDs) was observed, confirming HLSC growth, identity and functionality. For the expansion of HLSCs in the multi-plate bioreactor system Xpansion (XPN), the oxygen supply strategy was optimized due to a low k_La of 0.076 h⁻¹. The XPN bioreactor yielded a final mean cell density of 94 ± 8 × 10³ cells cm⁻², more than double that of the standard process in T-flasks. However, in the larger XPN50 device, HLSC density reached only 28 ± 0.9 × 10³ cells cm⁻², while the glucose consumption rate increased 8-fold. In a fully-controlled 2 L stirred-tank bioreactor (STR), HLSCs expanded at a comparable rate to the T-flask and XPN50 processes in a homogeneous microenvironment using advanced process analytical technology. Ultimately, the scale-up of HLSCs was successful using two different bioreactor systems, resulting in sufficient numbers of viable, functional and undifferentiated HLSCs for therapeutic applications. Full article