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The in vitro growth of a three‐dimensional human dermal replacement using a single‐pass perfusion system
Author(s) -
Halberstadt Craig R.,
Hardin Robert,
Bezverkov Kelly,
Snyder Dale,
Allen Lynn,
Landeen Lee
Publication year - 1994
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260430808
Subject(s) - bioreactor , fibronectin , perfusion , chemistry , ascorbic acid , biomedical engineering , matrix (chemical analysis) , glycosaminoglycan , in vitro , residence time distribution , biochemistry , biophysics , chromatography , extracellular matrix , food science , biology , medicine , mineralogy , inclusion (mineral) , organic chemistry
A human dermal replacement has been developed by seeding human neonatal dermal fibroblasts onto a biosorbable polyglactin (polyglycolide/polylactide) mesh and culturing in a bioreactor. The mesh provides the proper environment for the cells to attach, grow in a three‐dimensional array, and establish a tissue matrix over a 2‐ to 3‐week culture period. The dermal replacement has been characterized and found to contain a variety of naturally occurring dermal matrix proteins, including fibronectin, glycosaminoglycans, and collagen types I and III. To efficiently and reproducibly produce this dermal tissue equivalent, a closed, single‐pass perfusion system was developed and compared with a static process. In the single‐pas perfusion system, growth medium (containing ascorbic acid) was perfused around the 4 × 6 in. pieces of mesh at specific flow rates determined by nutrient consumption and waste production rates. The flow rates used for this system indicate that a diffusion‐limited regime exists with a mean residence time greater than 1 h for essential nutrients and factors. By controlling glucose concentrations in the system to a delta of 0.70 g/L from the inlet to the outlet of the bioreactor, it took 6 fewer days to grow a tissue similar to that produced by the static system.