High flow technique for harvesting mammalian cells

Chemistry: molecular biology and microbiology – Process of utilizing an enzyme or micro-organism to destroy... – Treating animal or plant material or micro-organism

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435 11, 435 12, C07G 1500, A01N 102

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059121637

ABSTRACT:
The invention is based in part on the observation that significant portions of porcine livers appear to remain intact after perfusion by standard methods, suggesting that the perfusion procedures employed do not result in complete enzymatic digestion. Recovery of cells is therefore substantially lower than would be possible if the organs were thoroughly digested. It is found that increased perfusion flow rate, occlusion of at least one major blood vessel leading out of the organ, increased enzymatic digestion time, and vigorous tissue dissociation techniques can be combined to afford a uniquely high yield of viable cells.

REFERENCES:
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Gerlach et al., "Comparison of Four Methods for Mass Hepatocyte Isolation from Pig and Human Livers", Transplantation 57(9): 1318-22 (1994).
Gerlach et al., "Nonenzymatic versus enzymatic hepatocyte isolation from pig livers for larger scale investigations of liver cell perfusion systems," Intl. J. Artificial Organs, 16(9):677-681, 1993.
Koebe et al., "Porcine Hepatocytes for Biohybrid Artifical Liver Devices: A Comparison of Hypothermic Storage Techniques," Artif. Organ, 20(11):1181-1190, 1996.
Koebe and Schildberg, "Isolation of porcine hepatocytes from slaughterhouse organs," Intl. J. Artificial Organs, 19(1):53-60, 1996.
Morsiani et al., "Automatic Liver Cell Processing Facilitates Large Scale Isolation and Purification of Porcine Hepatocytes," ASAIO Journal, 41:155-161, 1994.
Naik et al., "Isolation and Culture of Porcine Hepatocytes for Artificial Liver Support," Cell Transplantation, 5(1):107-115, 1996.
Seglen, "Preparation of Isolated Rat Liver Cells," Norsk Hydro's Institute for Cancer Research, Department of Tissue Culture, The Norwegian Raidum Hospital, Montebello, Oslo, Norway, 29-83, 1976.
Sielaff et al., "A Technique for Porcine Hepatocyte Harvest and Description of Differentiated Metabolic Functions in Static Culture," U. Minnesota Medical School, Coon Rapids, MN 1995.

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