title: Culture and function of electrofusion-derived clonal insulin-secreting cells immobilized on solid and macroporous microcarrier beads.
abstract: In view of the advantages of the bulk production of clonal pancreatic beta cells, an investigation was made of the growth and insulin secretory functions of an electrofusion-derived cell line (BRIN-BD11) immobilized on a solid microcarrier, cytodex-1 or a macroporous microcarrier, cultispher-G. For comparison, similar tests were performed using BRIN-BD11 cells present in single cell suspensions or allowed to form pseudoislets. Similar growth profiles were recorded for each microcarrier with densities of 4.4 x 10(5) +/- 0.3 cells/ml and 4.2 x 10(5) +/- 0.2 cells/ml achieved using cytodex-1 and cultispher-G, respectively. Cell viability began to decline on day 5 of culture. Insulin concentration in the culture medium reached a peak of 26 +/- 2.0 ng/ml and 24 +/- 2.2 ng/ml for cells grown on cytodex-1 and cultispher-G, respectively. Cells grown on both types of microcarrier showed a significant 1.5-1.8-fold acute insulin-secretory response to 16.7 mmol/l glucose. L-alanine (10 mmol/l) and L-arginine (10 mmol/l) also induced significant 3 4 fold increases of insulin release. BRIN-BD11 cells immobilized on cytodex-1 or cultispher-G out-performed single cell suspensions and pseudoislets in terms of insulin-secretory responses to glucose and amino acids. A 1.3-fold, 2.2-fold and 1.7-fold stimulation of insulin secretion was observed for glucose, L-alanine and L-arginine respectively in single cell suspensions. Corresponding increases for pseudoislets were 1.6-1.8-fold for L-alanine and L-arginine, with no significant response to glucose alone. These data indicate the utility of micro-carriers for the production of functioning clonal beta cells.
authors.affiliation: School of Biomedical Sciences, University of Ulster, Coleraine, Northern Ireland, UK.
authors.forename: M
authors.initials: M
authors.lastname: Hamid
authors.forename: J T
authors.initials: JT
authors.lastname: McCluskey
authors.forename: N H
authors.initials: NH
authors.lastname: McClenaghan
authors.forename: P R
authors.initials: PR
authors.lastname: Flatt
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division; D002459:Cell Fusion; D002460:Cell Line; D002470:Cell Survival; D002999:Clone Cells / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D003911:Dextrans / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D007328:Insulin / Q000096:biosynthesis / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism*; D008863:Microspheres; D051381:Rats
chemical_list: D000596:Amino Acids; D003470:Culture Media; D003911:Dextrans; D007328:Insulin; C052839:Cytodex; D005947:Glucose