Human proinsulin production in primary rat hepatocytes after retroviral vector gene transfer

Luca Falqui, Sabina Martinenghi, Cesare Berra, Lucilla Monti, Biagio E. Leone, Guido Pozza, Claudio Bordignon

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The development of autologous somatic cells, engineered for the synthesis and release of human insulin under physiological stimuli, would certainly represent a major breakthrough in the therapy of insulin-dependent diabetes mellitus. We generated a retroviral vector containing the human proinsulin cDNA and the gene coding for the human nerve growth factor receptor for quantitative analysis of transduced cells. Primary rat hepatocytes were selected as target cells because of the constitutive expression of the pancreatic β-cell glucose transporter GLUT-2 and the glycolitic enzyme glucokinase. Appropriate conditions for culture and retroviral transduction are described. The highest transduction efficiency, evaluated as percentage of LNGFr expressing cells was obtained by repeated infection cycles (40±10%). Human proinsulin accumulated in the culture medium of transduced rat hepatocytes (mean±SD): 18.1±7.9 (range 8.7-36.4) ng/24h/10 6 cells. Primary rat hepatocytes can be efficiently transduced by a retroviral vector and the de novo synthesis of human proinsulin can be induced. Primary cultured hepatocytes represent an useful model to test retroviral constructs engineered for the glucose-inducible expression of insulin under the control of liver-specific promoters.

Original languageEnglish
Pages (from-to)250-253
Number of pages4
JournalJournal of Molecular Medicine
Volume77
Issue number1
Publication statusPublished - 1999

Fingerprint

Proinsulin
Hepatocytes
Genes
Insulin
Glucokinase
Nerve Growth Factor Receptor
Facilitative Glucose Transport Proteins
Type 1 Diabetes Mellitus
Culture Media
Complementary DNA
Glucose
Liver
Enzymes
Infection

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Falqui, L., Martinenghi, S., Berra, C., Monti, L., Leone, B. E., Pozza, G., & Bordignon, C. (1999). Human proinsulin production in primary rat hepatocytes after retroviral vector gene transfer. Journal of Molecular Medicine, 77(1), 250-253.

Human proinsulin production in primary rat hepatocytes after retroviral vector gene transfer. / Falqui, Luca; Martinenghi, Sabina; Berra, Cesare; Monti, Lucilla; Leone, Biagio E.; Pozza, Guido; Bordignon, Claudio.

In: Journal of Molecular Medicine, Vol. 77, No. 1, 1999, p. 250-253.

Research output: Contribution to journalArticle

Falqui, L, Martinenghi, S, Berra, C, Monti, L, Leone, BE, Pozza, G & Bordignon, C 1999, 'Human proinsulin production in primary rat hepatocytes after retroviral vector gene transfer', Journal of Molecular Medicine, vol. 77, no. 1, pp. 250-253.
Falqui, Luca ; Martinenghi, Sabina ; Berra, Cesare ; Monti, Lucilla ; Leone, Biagio E. ; Pozza, Guido ; Bordignon, Claudio. / Human proinsulin production in primary rat hepatocytes after retroviral vector gene transfer. In: Journal of Molecular Medicine. 1999 ; Vol. 77, No. 1. pp. 250-253.
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