Breast cancer is the most common invasive cancer in women worldwide. Surgical removal of the breast tumor and subsequent reconstructive surgery can result in complications such as infection or necrosis of transplanted adipose tissue. Breast cancer recurrence is also a serious concern for patients. Our group has developed a tannic acid/collagen bead material that has the potential to be used as an injectable adipose tissue regenerative device to replace lipofilling. Tannic acid is a polyphenol with anticancer and antibiotic properties. The objective of this study was to establish the biocompatibility of an injectable tannic acid/collagen bead implant material in an in vivo rat model. The injection of the tannic acid-collagen type I bead device was minimally invasive. No symptoms of infection, tissue necrosis, or widespread chronic inflammation were observed. After 12 weeks, implants showed incorporation into native tissue with no fibrous encapsulation. Despite the presence of inflammatory cells in the remaining beads, fat tissue growth and collagen redistribution were observed within the beads over 12 weeks, showing incorporation within native subcutaneous tissue and indicating good biocompatibility and bioactivity of the implant. Our results demonstrate that the tannic acid/collagen bead scaffold has good biocompatibility and works as an adipocyte tissue regeneration and reconstructive device.
Breast cancer is the most commonly diagnosed cancer among women worldwide. Surgical removal of tumors is often necessary and many patients suffer complications due to subsequent breast reconstruction. A safe and effective breast reconstructive material is needed for patients recovering from surgical removal of small breast cancer tumors. Our lab has developed injectable collagen/tannic acid beads seeded with patient-derived preadipocytes for regeneration of healthy breast tissue in patients post-lumpectomy. Previous research indicates that the inclusion of tannic acid in the matrix imparts an anticancer property. This research seeks to determine the variables needed to control collagen/tannic acid bead diameter and seeded cell attachment, which are essential to proper bead implantation and function. We found that as tannic acid concentration increases within the beads, cell attachment decreases. Bead diameter is controlled by bead generator voltage, solution osmolality, the degree of cell attachment, and tannic acid concentrations. Higher voltages resulted in significant decrease in bead diameter. Collagen/tannic acid beads decreased in diameter when placed in solutions of increasing osmolality. Higher degrees of cell attachment across the surface of the beads were associated with a significant decrease in diameter. In beads made with high concentrations of tannic acid, bead diameter was found to decrease. Collagen/TA beads are a promising subdermal tissue regenerative matrix with anticancer activity as an alternative to simple lipofilling in breast reconstructive procedures. This study was conducted to better understand the properties of collagen/TA beads in order to improve injection efficacy and tissue regenerative activity.
Due to the favorable attributes of Chinese hamster ovary (CHO) cells for therapeutic proteins and antibodies biomanufacturing, companies generate proprietary cells with desirable phenotypes. One key attribute is the ability to stably express multi‐gram per liter titers in chemically defined media. Cell, media, and feed diversity has limited community efforts to translate knowledge. Moreover, academic, and nonprofit researchers generally cannot study “industrially relevant” CHO cells due to limited public availability, and the time and knowledge required to generate such cells. To address these issues, a university‐industrial consortium (Advanced Mammalian Biomanufacturing Innovation Center, AMBIC) has acquired two CHO “reference cell lines” from different lineages that express monoclonal antibodies. These reference cell lines have relevant production titers, key performance outcomes confirmed by multiple laboratories, and a detailed technology transfer protocol. In commercial media, titers over 2 g/L are reached. Fed‐batch cultivation data from shake flask and scaled‐down bioreactors is presented. Using productivity as the primary attribute, two academic sites aligned with tight reproducibility at each site. Further, a chemically defined media formulation was developed and evaluated in parallel to the commercial media. The goal of this work is to provide a universal, industrially relevant CHO culture platform to accelerate biomanufacturing innovation.
The accumulation of metabolic wastes in cell cultures can diminish product quality, reduce productivity, and trigger apoptosis. The limitation or removal of unintended waste products from Chinese hamster ovary (CHO) cell cultures has been attempted through multiple process and genetic engineering avenues with varied levels of success. One study demonstrated a simple method to reduce lactate and ammonia production in CHO cells with adaptation to extracellular lactate; however, the mechanism behind adaptation was not certain. To address this profound gap, this study characterizes the phenotype of a recombinant CHO K-1 cell line that was gradually adapted to moderate and high levels of extracellular lactate and examines the genomic content and role of extrachromosomal circular DNA (eccDNA) and gene expression on the adaptation process. More than 500 genes were observed on eccDNAs. Notably, more than 1000 genes were observed to be differentially expressed at different levels of lactate adaptation, while only 137 genes were found to be differentially expressed between unadapted cells and cells adapted to grow in high levels of lactate; this suggests stochastic switching as a potential stress adaptation mechanism in CHO cells. Further, these data suggest alanine biosynthesis as a potential stress-mitigation mechanism for excess lactate in CHO cells.
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