New Miniaturized Hollow-Fiber Bioreactor for in Vivo Like Cell Culture, Cell Expansion, and Production of Cell-Derived Products

Abstract

We have developed a miniaturized hollow-fiber bioreactor system for mammalian cell culture with a volume of 1 mL. Cell and medium compartments of the bioreactor are separated by a semipermeable membrane, and oxygenation of the cell compartment is accomplished using an oxygenation membrane. As a result of the geometry of the transparent housing, cells can be observed by microscopy during culture. The leukemic cell lines CCRF-CEM, HL-60, and REH were cultivated up to densities of 3.5 x 10(7)/mL without medium change or manipulation of the cells. As shown using CCRF-CEM cells, growth in the bioreactor was strongly influenced and could be controlled by the medium flow rate. As a consequence, consumption of glucose and generation of lactate varied with flow rate. Depending on the molecular size cutoff of the membranes used, added growth factors such as GM-CSF, as well as factors secreted from the cells, are retained in the cell compartment for up to 1 week. This new miniaturized hollow-fiber bioreactor offers advantages in tissue engineering by continuous nutrient supply for cells in high density, retention of added or autocrine produced factors, and undisturbed long-term culture in a closed system.

Single-cell-bioreactors as end of miniaturization approaches in biotechnology: progresses with characterised bioreactors and a glance into the future

“…These micro-compartments can be established via creation of a micro-membrane using wide-spread encapsulation techniques based on alginate and related matrices forming droplets with a co-acervate membrane [25,59,60]. Another way of establishing a microenvironment is a complete reactor re-design, incorporating smallest reactor compartments via hollow fibres or membranes in a flat arrangement [25].…”

“…Another way of establishing a microenvironment is a complete reactor re-design, incorporating smallest reactor compartments via hollow fibres or membranes in a flat arrangement [25].…”

“…Another well-established cultivation system employing smallest compartments is subsumable under the heading of 'hollow fibre reactor systems' or 'membrane reactors' [25]. These systems are an adoption of the spherical microcapsule arrangement to a linear hollow fibre or flat membrane arrangement with defined membrane properties.…”

Magnetic Nanoparticles for Tissue Engineering

“…In addition, culture volume inevitably becomes large, since there are numerous nonadherent cells such as hematopoietic cells that must be diluted and removed by washing and changes in medium, resulting in low-density culture of MSCs. Under such low-density culture conditions, MSCs do not proliferate immediately, and require much time to develop into colonies as growth factors, including autocrine [21][22][23][24], paracrine [24], and juxtacrine [25] factors, play an important role in cell growth. Therefore, we developed a new method for producing high-density culture, using magnetic nanoparticles to promote expansion of MSCs.…”

Collection of in vivo-like liver cell secretome with alternative sample enrichment method using a hollow fiber bioreactor culture system combined with tangential flow filtration for secretomics analysis

“…The HFB culture system has several advantages over traditional dish or flask culture techniques. First, the HFB culture system can support the growth of a large number of cells in a small volume and permits a continuous process of cell culture that approaches tissue-like cell densities [16][17][18]. To maintain normal functions in the hepatocyte, an adequate mass of hepatocyte is needed, which depends on cell density [16].…”