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Isolation, Expansion, and Adipogenic Induction of CD34+CD31+ Endothelial Cells from Human Omental and Subcutaneous Adipose Tissue
Published on: July 17, 2018
Adipose Tissue is A Better Source of Immature Non-Hematopoietic Cells than Bone Marrow
Juri Fujimura1, Hidemitsu Sugihara2, Yoshitaka Fukunaga3
1Department of Pediatrics, Nippon Medical School, Tokyo, Japan ; Department of Pharmacology, Nippon Medical School, Tokyo, Japan.
Adipose tissue (AT) is an alternative source of the adult stem cells that can also be harvested from bone marrow (BM). Cultured AT-derived stem cells (ASCs) have been well characterized by many groups. However, non-cultured ASCs remain to be characterized. Hoechst 33342 dye efflux is a characteristic that is common to stem cells, as well as chemotherapy-resistant cancer cells. Thus, we compared the Hoechst 33342-stained side population (SP) cells in murine adipose-tissue (AT-SP cells) to the SP cells from murine bone marrow (BM-SP cells). The AT-SP cells were detected much more frequently in the 22 AT samples that were tested (0.42∼6.00%, mean 2.57%) than the BM-SP cells were detected in the 6 BM samples (0.02∼0.36%, mean 0.12%). After Hoechst staining, SP cells were analyzed by fluorescence-activated cell sorting (FACS) and electron micrograms. FACS analysis revealed that the AT-SP cells were CD44-, CD45-, CD45R+, Sca-1± and c-kit-, while the BM-SP cells were CD44-, CD45±, CD45R-, Sca-1+ and c-kit+. This indicates that the AT-SP cells differ phenotypically from the BM-SP cells. Electron microscopic analysis revealed that the AT-SP cells are small cells with a diameter of about 5 um. Some of the BM-SP cells had granules, similar to eosinophils or basophils, whereas the AT-SP cells had fewer organelles and a higher N/C ratio than the BM-SP cells. This suggests that the AT-SP cells are considerably more immature than the BM-SP cells. Thus, it appears that AT is a better source of immature non-hematopoietic cells than BM.
Adipose tissue (AT) is an alternative source of the adult stem cells that can also be harvested from bone marrow (BM). Cultured AT-derived stem cells (ASCs) have been well characterized by many groups. However, non-cultured ASCs remain to be characterized. Hoechst 33342 dye efflux is a characteristic that is common to stem cells, as well as chemotherapy-resistant cancer cells. Thus, we compared the Hoechst 33342-stained side population (SP) cells in murine adipose-tissue (AT-SP cells) to the SP cells from murine bone marrow (BM-SP cells). The AT-SP cells were detected much more frequently in the 22 AT samples that were tested (0.42∼6.00%, mean 2.57%) than the BM-SP cells were detected in the 6 BM samples (0.02∼0.36%, mean 0.12%). After Hoechst staining, SP cells were analyzed by fluorescence-activated cell sorting (FACS) and electron micrograms. FACS analysis revealed that the AT-SP cells were CD44-, CD45-, CD45R+, Sca-1± and c-kit-, while the BM-SP cells were CD44-, CD45±, CD45R-, Sca-1+ and c-kit+. This indicates that the AT-SP cells differ phenotypically from the BM-SP cells. Electron microscopic analysis revealed that the AT-SP cells are small cells with a diameter of about 5 um. Some of the BM-SP cells had granules, similar to eosinophils or basophils, whereas the AT-SP cells had fewer organelles and a higher N/C ratio than the BM-SP cells. This suggests that the AT-SP cells are considerably more immature than the BM-SP cells. Thus, it appears that AT is a better source of immature non-hematopoietic cells than BM.
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