期刊
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY
卷 61, 期 10, 页码 719-730出版社
SAGE PUBLICATIONS LTD
DOI: 10.1369/0022155413502055
关键词
adipocyte; mesenchymal stem cell; basement membrane; differentiation; laminin; integrin
类别
资金
- Nona and Kullervo Vare Foundation
- Paivikki and Sakari Sohlberg Foundation
- Sigrid Juselius Foundation [290312]
- HUS-evo grants [131210]
- TBDP PhD Graduate School [031111]
- Danish Council for Strategic Research [4270289]
- Invalid Foundation [9310/407]
- Magnus Ehrnrooth Foundation
- Perklen Foundation
- Liv och Halsa Foundation
- Orion-Farmos Tutkimussaatio
- Finska Lakaresallskapet [131210]
The aim was to study laminin (LM) synthesis, integration, and deposition into the basement membrane (BM) during adipogenesis. Human bone marrow-derived mesenchymal stromal cells (MSCs) were induced along the adipogenic lineage. LM chain mRNA and protein levels were followed using quantitative real-time polymerase chain reaction (qRT-PCR), immunofluorescence (IF) staining, transmission electron microscopy (TEM), and immunoprecipitation. MSCs produced low levels of LM mRNAs but were not surrounded by BM in IF and TEM imaging. LM-4, LM-1, and LM-1 mRNAs increased during adipogenesis 3.9-, 5.8-, and 2.8-fold by day 28. LM-411 was immunoprecipitated from the ECM of the differentiated cells. Immunostaining suggested deposition of LM-411 and some LM-421. BM build-up was probably organized in part by integrin (Int) (61). At day 28, TEM images revealed BM-like structures around fat droplet-containing cells. The first signs of BM formation and Int (61) were seen using IF imaging at day 14. Laminin-411 and Int (61) were expressed in vivo in mature human subcutaneous fat tissue. Undifferentiated human MSCs did not organize LM subunits into BM, whereas LM-411 and some LM-421 are precipitated in the BM around adipocytes. This is the first demonstration of LM-411 precipitation during hMSC adipogenesis around adipocytes as a structural scaffold and Int-regulated signaling element.
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