Nestin-positive progenitor cells isolated from human fetal pancreas have phenotypic markers identical to mesenchymal stem cells.

AIM To isolate nestin-positive progenitor cells from human fetal pancreas and to detect their surface markers and their capability of proliferation and differentiation into pancreatic islet endocrine cells in vitro. METHODS Islet-like cell clusters (ICCs) were isolated from human fetal pancreas by using collagenase digestion. The free-floating ICCs were handpicked and cultured in a new dish. After the ICCs developed into monolayer epithelium-like cells, they were passaged and induced for differentiation. Reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence stain, fluorescence-activated cell sorting (FACS) and radioimmunoassay (RIA) were used to detect the expression of cell markers. RESULTS (1) The monolayer epithelium-like cells had highly proliferative potential and could be passaged more than 16 times in vitro; (2) RT-PCR analysis and immunofluorescence stain showed that these cells expressed both nestin and ABCG2, two of stem cell markers; (3) FACS analysis revealed that CD44, CD90 and CD147 were positive, whereas CD34, CD38, CD45, CD71, CD117, CD133 and HLA-DR were negative on the nestin-positive cells; (4) RT-PCR analysis showed that the mRNA expression of insulin, glucagon and pancreatic-duodenal homeobox gene-1 was detected, whereas the expression of nestin and neurogenin 3 disappeared in these cells treated with serum-free media supplemented with the cocktail of growth factors. Furthermore, the intra-cellular insulin content was detected by RIA after the induction culture. CONCLUSION Nestin-positive cells isolated from human fetal pancreas possess the characteristics of pancreatic progenitor cells since they have highly proliferative potential and the capability of differentiation into insulin-producing cells in vitro. Interestingly, the nestin-positive pancreatic progenitor cells share many phenotypic markers with mesenchymal stem cells derived from bone marrow.

[1]  Ana D. Lopez,et al.  Characterization and isolation of promoter-defined nestin-positive cells from the human fetal pancreas. , 2003, Diabetes.

[2]  Donna M. Martin,et al.  Nestin-lineage cells contribute to the microvasculature but not endocrine cells of the islet. , 2003, Diabetes.

[3]  O. Korsgren,et al.  Characterization of endocrine progenitor cells and critical factors for their differentiation in human adult pancreatic cell culture. , 2003, Diabetes.

[4]  D. Kotton,et al.  Side population cells and Bcrp1 expression in lung. , 2003, American journal of physiology. Lung cellular and molecular physiology.

[5]  Xue-Ming Tang,et al.  Phenotypic Determination and Characterization of Nestin-Positive Precursors Derived from Human Fetal Pancreas , 2003, Laboratory Investigation.

[6]  Gabriela Kania,et al.  Expression of Pax4 in embryonic stem cells promotes differentiation of nestin-positive progenitor and insulin-producing cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Y. Romanov,et al.  Searching for Alternative Sources of Postnatal Human Mesenchymal Stem Cells: Candidate MSC‐Like Cells from Umbilical Cord , 2003, Stem cells.

[8]  M. Hussain,et al.  In vivo derivation of glucose-competent pancreatic endocrine cells from bone marrow without evidence of cell fusion. , 2003, The Journal of clinical investigation.

[9]  Seung K. Kim,et al.  Growth inhibitors promote differentiation of insulin-producing tissue from embryonic stem cells , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[10]  M. Rudnicki,et al.  Side population cells from diverse adult tissues are capable of in vitro hematopoietic differentiation. , 2002, Experimental hematology.

[11]  M. Rudnicki,et al.  The post‐natal heart contains a myocardial stem cell population , 2002, FEBS letters.

[12]  C. Verfaillie,et al.  Multipotent progenitor cells can be isolated from postnatal murine bone marrow, muscle, and brain. , 2002, Experimental hematology.

[13]  R. Schwartz,et al.  Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells. , 2002, The Journal of clinical investigation.

[14]  K. Seuwen,et al.  Stem cell characteristics of human trabecular bone-derived cells. , 2002, Bone.

[15]  N. Fisk,et al.  Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. , 2001, Blood.

[16]  H. Nakauchi,et al.  The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype , 2001, Nature Medicine.

[17]  R. McKay,et al.  Differentiation of Embryonic Stem Cells to Insulin-Secreting Structures Similar to Pancreatic Islets , 2001, Science.

[18]  J. Habener,et al.  Multipotential nestin-positive stem cells isolated from adult pancreatic islets differentiate ex vivo into pancreatic endocrine, exocrine, and hepatic phenotypes. , 2001, Diabetes.

[19]  G. Fantuzzi,et al.  Interleukin-18 mRNA, but not interleukin-18 receptor mRNA, is constitutively expressed in islet beta-cells and up-regulated by interferon-gamma. , 2000, European cytokine network.

[20]  P. Quesenberry,et al.  Rapid Communication Characterization of Neurosphere Cell Phenotypes by Flow Cytometry , 2000 .

[21]  M. Pittenger,et al.  Human mesenchymal stem cells: progenitor cells for cartilage, bone, fat and stroma. , 2000, Current topics in microbiology and immunology.

[22]  Ames,et al.  Islet Transplantation in Seven Patients with Type 1 Diabetes Mellitus Using a Glucocorticoid-Free Immunosuppressive Regimen , 2000 .

[23]  M. Goodell,et al.  Hematopoietic potential of stem cells isolated from murine skeletal muscle. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[24]  R. Mulligan,et al.  Dystrophin expression in the mdx mouse restored by stem cell transplantation , 1999, Nature.

[25]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[26]  R. Johnson,et al.  Dye efflux studies suggest that hematopoietic stem cells expressing low or undetectable levels of CD34 antigen exist in multiple species , 1997, Nature Medicine.

[27]  J. Rubin,et al.  Regulation of Proliferation and Differentiation of Human Fetal Pancreatic Islet Cells by Extracellular Matrix, Hepatocyte Growth Factor, and Cell-Cell Contact , 1996, Diabetes.

[28]  A. S. Conner,et al.  Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo , 1996, The Journal of experimental medicine.

[29]  C. Ricordi,et al.  Nicotinamide is a potent inducer of endocrine differentiation in cultured human fetal pancreatic cells. , 1993, The Journal of clinical investigation.

[30]  R. McKay,et al.  Characterization of the human nestin gene reveals a close evolutionary relationship to neurofilaments. , 1992, Journal of cell science.

[31]  R. McKay,et al.  CNS stem cells express a new class of intermediate filament protein , 1990, Cell.