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Isolation and identification of mesenchymal stem cells from human fetal pancreas
Ying Hu1, Lianming Liao, Qiuying Wang
1State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Tianjin, People's Republic of China.
Abstract:
Mesenchymal stem cells (MSCs) have been cultured from many sources, including bone marrow and liver. To further support our hypothesis that MSCs exist in most postnatal tissues, we isolated a clonogenic, multipotent, rapidly proliferating population of cells from a fetal pancreas and termed them "pancreas-derived mesenchymal stem cells" (PMSCs). They withstood being passaged as many as 30 times without sustaining significant structural changes. In this study, we showed that PMSCs are positive for CD44, CD29, and CDI3 but negative for CD34 and HLA-DR and that they stained with collagen I and III but not with von Willebrand factor antibody. During the log phase of growth, PMSCs proliferated, doubling in population in about 30 hours. Cell-cycle analysis showed that more than 90% of cells were in the G0 and G1 phases, whereas a small subpopulation of cells were actively engaged in proliferation (S + G2 + M = 3.55%). Under differentiation culture conditions, PMSCs differentiated into cells of osteogenic, chondrogenic, and adipogenic lineages. These results demonstrate that PMSCs can be isolated from human fetal pancreas by means of their adherent ability and that they are capable of self-renewal, propagation, and multipotent differentiation.
Insights
Pancreas-derived mesenchymal stem cells (PMSCs) were isolated and characterized. These multipotent stem cells demonstrate self-renewal and differentiation capabilities, supporting their presence in postnatal tissues.
Area of Science:
- Stem Cell Biology
- Developmental Biology
- Regenerative Medicine
Background:
- Mesenchymal stem cells (MSCs) are found in various postnatal tissues.
- The existence of MSCs in most postnatal tissues is hypothesized.
- Pancreatic tissue is a potential source for novel stem cell populations.
Purpose of the Study:
- To isolate and characterize pancreas-derived mesenchymal stem cells (PMSCs).
- To investigate the self-renewal, proliferation, and multipotent differentiation potential of PMSCs.
- To support the hypothesis of MSCs existing in diverse postnatal tissues.
Main Methods:
- Isolation of clonogenic cells from human fetal pancreas.
- Culture and passaging of cells up to 30 times.
- Immunophenotyping using specific cell surface markers (CD44, CD29, CD13, CD34, HLA-DR).
- Assessment of differentiation into osteogenic, chondrogenic, and adipogenic lineages.
- Cell-cycle analysis to determine proliferation rates.
Main Results:
- Successfully isolated and cultured pancreas-derived mesenchymal stem cells (PMSCs).
- PMSCs exhibited positive staining for CD44, CD29, CD13 and collagen I/III, but negative for CD34 and HLA-DR.
- Cells demonstrated robust proliferation (doubling time ~30 hours) with >90% in G0/G1 phase.
- PMSCs underwent successful differentiation into osteogenic, chondrogenic, and adipogenic lineages.
- Cells maintained structural integrity over multiple passages.
Conclusions:
- Pancreas-derived mesenchymal stem cells (PMSCs) can be isolated from human fetal pancreas.
- PMSCs possess characteristics of mesenchymal stem cells, including self-renewal and multipotency.
- These findings support the hypothesis that mesenchymal stem cells are present in most postnatal tissues.