Nestin-positive progenitor cells derived from adult human pancreatic islets of Langerhans contain side population (SP) cells defined by expression of the ABCG2 (BCRP1) ATP-binding cassette transporter

doi:10.1016/S0006-291X(02)00275-9

Biochemical and Biophysical Research Communications

Volume 293, Issue 2, 3 May 2002, Pages 670-674

https://doi.org/10.1016/S0006-291X(02)00275-9 Get rights and content

Abstract

The disease diabetes mellitus arises as a consequence of a failure of the β-cells in the islets of Langerhans of the pancreas to produce insulin in the amounts required to meet the needs of the body. Whole pancreas or islet transplants in patients with severe diabetes effectively restore insulin production. A lack of availability of donor pancreata requires the development of alternative sources of islets such as the ex vivo culture and differentiation of stem/progenitor cells. Earlier we discovered multipotential progenitor cells in islets isolated from adult human pancreata that express the neural stem cell marker nestin: nestin-positive islet-derived progenitor cells (NIPs). Recently it was shown that the exclusion of the Hoechst 33342 dye, which defines the pluripotential side population (SP) of hematopoietic stem cells, is mediated by the ATP-binding cassette transporter, ABCG2. Here we report that the human islet-derived NIPs contain a substantial subpopulation of SP cells that co-express ABCG2, MDR1, and nestin. Thus NIPs may be a potential source of adult pluripotential stem/progenitor cells useful for the production of islet tissue for transplantation into diabetic subjects.

Section snippets

Materials and methods

Isolation and culture of NIPs. Human pancreatic islets were obtained from the islet distribution programs of the Cell Transplant Center, Diabetes Research Institute, University of Miami School of Medicine (Miami, FL), and the Juvenile Diabetes Research Foundation Center for Islet Transplantation, Harvard Medical School (Boston, MA). NIPs were isolated as previously described [3]. Briefly, human pancreatic islets were handpicked, incubated at 37 °C at 5% CO₂ in concanavalin A (ConA) coated

The ATP-dependent transporter ABCG2 is expressed in nestin-positive cells derived from pancreatic islets

NIPs were isolated from the outgrowth of human pancreatic islets and cultured until they reached confluence. RT-PCR demonstrated the expression of ABCG2 (Fig. 1A). The RT-PCR-generated DNA product was sequenced and shown to be identical (586 bp) with that of the human ABCG2 (GenBank Accession No. XM-032424) (data not shown). Southern blot hybridization with a cloned ABCG2 probe confirmed the amplification of the correct cDNA (Fig. 1A).

Cultured cells (1.5–2%) have a side population phenotype in the Hoechst 33342 dye exclusion assay

To investigate whether the expression of ABCG2 lends a side

Discussion

We have shown that nestin-positive cells derived from human pancreatic islets contain 1.5–2% of SP cells, which express ABCG2 and nestin at high levels compared to non-SP control cells. The correlation of ABCG2 expression with the SP phenotype in our experiments confirms the finding that ABCG2 activity is a major component of the SP dye efflux, recently reported by several groups [10], [11], [12]. To our knowledge the expression of nestin, originally found in neural stem/progenitor cells [4],

Acknowledgements

We thank Maris Handley at the Dana Farber Cancer Center for the help with FACS sorting, Julia Lin for the expert experimental assistance and Townley Budde for the help in the preparation of the manuscript. We are grateful for the provision of human islets by the University of Miami and Harvard University Centers for Islet Transplantation supported by the Juvenile Diabetes Research Foundation. The studies were supported in part by the USPHS Grant DK R21 55365. J.F.H. is an investigator with the

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Nestin-positive progenitor cells derived from adult human pancreatic islets of Langerhans contain side population (SP) cells defined by expression of the ABCG2 (BCRP1) ATP-binding cassette transporter

Abstract

Section snippets

Materials and methods

The ATP-dependent transporter ABCG2 is expressed in nestin-positive cells derived from pancreatic islets

Cultured cells (1.5–2%) have a side population phenotype in the Hoechst 33342 dye exclusion assay

Discussion

Acknowledgements

Cell

Neuron

Blood

Blood

Blood

Blood

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Blood

Neuron

Blood

Cell

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