Evaluating prostate cancer cell culturing methods: a comparison of cell morphologies and metabolic activity.
LNCaP prostate cancer cells were grown under four unique cultivation conditions. Two types of bioreactor systems were used to observe the influence of low-shear culture conditions allowing for three-dimensional growth: a) a perfusion rotating wall vessel (RWV) bioreactor; and b) a high aspect ratio vessel (HARV) RWV bioreactor, with periodic medium exchanges (fed-batch). In addition, two growth methods utilized tissue culture flasks (TCFs): a) unaltered or conventional TCFs; and b) poly(2-hydroxyethyl methacrylate) [poly(HEMA)] coated TCFs, to inhibit cell attachment. Comparisons were drawn based on qualitative observation of cell morphology and quantitative metabolic data. Similar cellular metabolism was demonstrated for cells grown under each condition. The degree of aggregation, however, varies considerably. Spherical shaped aggregates with diameters of 1 to 3 mm were produced when cells were grown within the perfusion-RWV bioreactor. All other growth conditions produced irregular shaped aggregates of various sizes. Quantitative results demonstrated the expected glucose utilization concomitant with lactate accumulation. Immunohistochemical evaluations were unremarkable for all four cultivation conditions. Results demonstrate that use of the perfusion-RWV bioreactor is advantageous in obtaining spherical aggregates, while grown in a controlled environment.
Winkenwerder
Jennifer J
JJ
Department of Chemistry, The University of Iowa, Iowa City, IA 52242, USA.
Palechek
Patricia L
PL
Reece
Julie S
JS
Saarinen
Mark A
MA
Arnold
Mark A
MA
Cohen
Michael B
MB
Murhammer
David W
DW
D019149:Bioreactors; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion; D002449:Cell Aggregation; D046508:Culture Techniques / Q000379:methods*; D005947:Glucose / Q000378:metabolism; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007150:Immunohistochemistry; D019344:Lactic Acid / Q000378:metabolism; D008297:Male; D011102:Polyhydroxyethyl Methacrylate / Q000737:chemistry; D017430:Prostate-Specific Antigen / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology*; D014407:Tumor Cells, Cultured
D015820:Cadherins; D018960:Hyaluronan Receptors; D011102:Polyhydroxyethyl Methacrylate; D019344:Lactic Acid; D017430:Prostate-Specific Antigen; D005947:Glucose