Carbone A, Valente M, Annacontini L, Castellani S, Di Gioia S, Parisi D, Rucci M, Belgiovine G, Colombo C, Di Benedetto A, Mori G, Lo Muzio L, Maiorella A, Portincasa A, Conese M
Fondazione IRCCS Ca Granda Ospedale Maggiore Policlinico, Medical Genetics Laboratory, Milan, Italy; Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy.
Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy.
J Biol Regul Homeost Agents. 2016 Jan-Mar;30(1):111-22.
The potential of adipose-derived mesenchymal stromal (stem) cells (ADSCs) to differentiate into either osteoblasts or chondrocytes is controversial. In this study we investigated the multicapacity potential of ADSCs to differentiate towards adipocyte, osteoblast, and chondrocyte lineages when cells are seeded onto plastic in comparison with incubation with conditioned media (CM) obtained from differentiated cell types.ADSCs, obtained from liposuctions, were characterized for mesenchymal and hematopoietic markers by cytofluorimetry. Their differentiation capacity towards adipocytes, osteoblasts, and chondrocytes was investigated by histochemistry methods (Oil-Red-O staining, Safranin O and Alizarin Red staining, respectively). Dental pulp stem cells (DPSCs) and dedifferentiated auricle derived-chondrocytes were differentiated towards osteoblastic and chondrocytic lineages respectively, and the CM obtained from these cultures was used to induce differentiation of ADSCs. ADSCs were positive for mesenchymal markers (CD29, CD105, CD73, CD44), but not for hematopoietic lineage markers (CD14, CD34, CD45) and this behavior was conserved from the isolation up to the fifth passage. While ADSCs were readily differentiated in adipocytes, they were not towards chondrocytes and osteoblastic lineages, a behavior different from that of bone marrow-derived MSCs that differentiated into the three lineages at two weeks post-induction. Only ADSCs treated with CM from cultured chondrocytes and DPSCs, produced glycosaminoglycans and mineralized matrix. These results indicate that ADSCs need growth/morphogenic factor supplementation from the tissue environment to be appropriately differentiated to mesodermic lineages.
脂肪来源的间充质基质(干)细胞(ADSCs)分化为成骨细胞或软骨细胞的潜能存在争议。在本研究中,我们调查了与接种到塑料培养板上相比,将ADSCs接种到从分化细胞类型获得的条件培养基(CM)中培养时,其向脂肪细胞、成骨细胞和软骨细胞谱系分化的多能潜能。通过吸脂获得的ADSCs,用细胞荧光分析法对间充质和造血标志物进行表征。通过组织化学方法(分别为油红O染色、番红O染色和茜素红染色)研究其向脂肪细胞、成骨细胞和软骨细胞的分化能力。牙髓干细胞(DPSCs)和去分化的耳廓软骨细胞分别向成骨细胞和软骨细胞谱系分化,从这些培养物中获得的CM用于诱导ADSCs分化。ADSCs对间充质标志物(CD29、CD105、CD73、CD44)呈阳性,但对造血谱系标志物(CD14、CD34、CD45)呈阴性,并且从分离到第五代这种特性都保持不变。虽然ADSCs很容易分化为脂肪细胞,但它们不能分化为软骨细胞和成骨细胞谱系,这一行为与骨髓来源的间充质干细胞不同,后者在诱导后两周可分化为这三种谱系。只有用培养的软骨细胞和DPSCs的CM处理的ADSCs才能产生糖胺聚糖和矿化基质。这些结果表明,ADSCs需要从组织环境中补充生长/形态发生因子才能适当地分化为中胚层谱系。