In vitro proliferation of cells derived from adult human beta-cells revealed by cell-lineage tracing

Diabetes. 2008 Jun;57(6):1575-83. doi: 10.2337/db07-1283. Epub 2008 Mar 3.


Objective: Expansion of insulin-producing beta-cells from adult human islets could alleviate donor shortage for cell-replacement therapy of diabetes. A major obstacle to development of effective expansion protocols is the rapid loss of beta-cell markers in the cultured cells. Here, we report a genetic cell-lineage tracing approach for following the fate of cultured beta-cells.

Research design and methods: Cells dissociated from isolated human islets were infected with two lentiviruses, one expressing Cre recombinase under control of the insulin promoter and the other, a reporter cassette with the structure cytomegalovirus promoter-loxP-DsRed2-loxP-eGFP.

Results: Beta-cells were efficiently and specifically labeled by the dual virus system. Label(+), insulin(-) cells derived from beta-cells were shown to proliferate for a maximum of 16 population doublings, with an approximate doubling time of 7 days. Isolated labeled cells could be expanded in the absence of other pancreas cell types if provided with medium conditioned by pancreatic non-beta-cells. Analysis of mouse islet cells by the same method revealed a much lower proliferation of labeled cells under similar culture conditions.

Conclusions: Our findings provide direct evidence for survival and dedifferentiation of cultured adult human beta-cells and demonstrate that the dedifferentiated cells significantly proliferate in vitro. The findings confirm the difference between mouse and human beta-cell proliferation under our culture conditions. These findings demonstrate the feasibility of cell-specific labeling of cultured primary human cells using a genetic recombination approach that was previously restricted to transgenic animals.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Cell Culture Techniques / methods
  • Cell Differentiation
  • Cell Division
  • Cell Survival
  • Cells, Cultured
  • Humans
  • Insulin-Secreting Cells / cytology*
  • Insulin-Secreting Cells / transplantation
  • Insulin-Secreting Cells / virology
  • Integrases / genetics
  • Integrases / metabolism
  • Kinetics
  • Lentivirus / enzymology
  • Lentivirus / genetics
  • Lentivirus / physiology
  • Polymerase Chain Reaction


  • Cre recombinase
  • Integrases