title: Differentiation and expansion of beta cell mass in porcine neonatal pancreatic cell clusters transplanted into nude mice.
abstract: Neonatal porcine pancreas has considerable capacity for growth and differentiation, making it an attractive potential source of islet tissue for xenotransplantation. Pancreases from 1-3-day-old newborn pigs were digested with collagenase and cultured for 8 days. The resulting cellular aggregates are called porcine neonatal pancreatic cell clusters (NPCCs). The mean yield of NPCCs from a newborn pig was 28,200 +/- 1700 islet equivalents. Cytokeratin 7 (CK7) was used as a marker for the immunostaining of pancreatic duct cells. In neonatal pancreas, 18% of the insulin-positive cells co-stained for CK7, thus being protodifferentiated. NPCCs also contained protodifferentiated cells; insulin/PP and insulin/somatostatin co-stained cells were more common than insulin/glucagon cells. Between 1 and 8 days of culture, the DNA content of the NPCCs fell to 16% and the insulin content to 33% of the starting value, mainly due to the preferential loss of exocrine cells. Transplantation of 2000 or 4000 NPCCs into diabetic nude mice typically normalized glucose values in 10-20 weeks. Mice with successful grafts had lower fasting blood glucose levels than normal mice and accelerated glucose clearance after an i.p. glucose load. The starting NPCCs consisted of 17% insulin-staining cells, but the grafts of mice with reversed diabetes consisted of 94% beta cells, with some co-stained for CK7, indicating that the grafts still contained immature cells. The mass of insulin-producing cells rose from 0.22 +/- 0.08 mg 1 week after transplantation to 4.34 +/- 0.27 mg in mice sacrificed at 27-35 weeks. In summary, NPCCs contain mostly islet precursor cells, which when transplanted into nude mice undergo striking differentiation and beta cell expansion.
authors.affiliation: Research Division, Joslin Diabetes Center, Boston, MA 02215, USA.
authors.forename: K H
authors.initials: KH
authors.lastname: Yoon
authors.forename: R R
authors.initials: RR
authors.lastname: Quickel
authors.forename: K
authors.initials: K
authors.lastname: Tatarkiewicz
authors.forename: T R
authors.initials: TR
authors.lastname: Ulrich
authors.forename: J
authors.initials: J
authors.lastname: Hollister-Lock
authors.forename: N
authors.initials: N
authors.lastname: Trivedi
authors.forename: S
authors.initials: S
authors.lastname: Bonner-Weir
authors.forename: G C
authors.initials: GC
authors.lastname: Weir
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001786:Blood Glucose / Q000378:metabolism; D002454:Cell Differentiation; D002455:Cell Division; D004247:DNA / Q000378:metabolism; D005951:Glucose Tolerance Test; D007150:Immunohistochemistry; D007328:Insulin / Q000097:blood / Q000378:metabolism; D016381:Islets of Langerhans Transplantation / Q000473:pathology* / Q000502:physiology; D053552:Keratin-7; D007633:Keratins / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D010191:Pancreatic Polypeptide / Q000378:metabolism; D013004:Somatostatin / Q000378:metabolism; D014183:Transplantation, Heterologous
chemical_list: D001786:Blood Glucose; D007328:Insulin; D053552:Keratin-7; C507496:Krt7 protein, mouse; D013004:Somatostatin; D010191:Pancreatic Polypeptide; D007633:Keratins; D004247:DNA