Flow cytometry and two-dimensional electrophoresis (2-DE) for system evaluation of long term continuous perfused animal cell cultures in macroporous beads

Abstract

Immobilization of r-CHO cells at high density using macroporous polyethylene carriers in a modular fluidized bed reactor is demonstrated. Specific growth rates of the cells are measured by incorporation of BrdU. At a cell density of about 10(8) cells/ml a stable growth rate of 0.004 h-1 was established. Total release of proteins into the culture supernatant during protein-free perfusion was analyzed by 2-DE in various phases of the long-term culture showing very similar patterns indicating a constant pattern of gene expression.

Cell growth on electrospun nanofiber mats from polyacrylonitrile (PAN) blends

“…As shown in Section 9.1.2.1, it is important to provide a constant environment for the production of cells by using perfusion technology (292).…”

hTERT alone immortalizes epithelial cells of renal proximal tubules without changing their functional characteristics

“…Determination of cell proliferation was done as described previously (41). After growth in medium supplemented with 50 M bromodeoxyuridine (BrdU) for 24 h, cells were stained with Hoechst 33258 and analyzed by flow cytometry (FACS Vantage; Becton Dickinson, Franklin Lakes, NJ).…”

High-density cultivation of insect cells and production of recombinant baculovirus using a novel oscillating bioreactor

“…This is often encountered because the processes associated with immobilized cells are often characterized by low specific growth rates (Reiter et al 1992). The repeatedbatch operation in this experiment was adequate to support high cell yield, but might be sub-optimal for fast cell growth.…”