Raman microspectroscopy reveals unsaturation heterogeneity at the lipid droplet level and validates an in vitro model of bone marrow adipocyte subtypes
Bone marrow adipocytes (BMAds) constitute the most abundant stromal component of adult human bone marrow. Two subtypes of BMAds have been described, the more labile regulated adipocytes (rBMAds) and the more stable constitutive adipocytes (cBMAds), which develop earlier in life and are more resilien...
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Frontiers Media S.A.
2022-11-01
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| Series: | Frontiers in Endocrinology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fendo.2022.1001210/full |
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| author | Josefine Tratwal Guillaume Falgayrac Alexandrine During Nicolas Bertheaume Charles Bataclan Daniel N. Tavakol Vasco Campos Ludovic Duponchel George Q. Daley Guillaume Penel Christophe Chauveau Olaia Naveiras Olaia Naveiras Olaia Naveiras |
| author_facet | Josefine Tratwal Guillaume Falgayrac Alexandrine During Nicolas Bertheaume Charles Bataclan Daniel N. Tavakol Vasco Campos Ludovic Duponchel George Q. Daley Guillaume Penel Christophe Chauveau Olaia Naveiras Olaia Naveiras Olaia Naveiras |
| author_sort | Josefine Tratwal |
| collection | DOAJ |
| description | Bone marrow adipocytes (BMAds) constitute the most abundant stromal component of adult human bone marrow. Two subtypes of BMAds have been described, the more labile regulated adipocytes (rBMAds) and the more stable constitutive adipocytes (cBMAds), which develop earlier in life and are more resilient to environmental and metabolic disruptions. In vivo, rBMAds are enriched in saturated fatty acids, contain smaller lipid droplets (LDs) and more readily provide hematopoietic support than their cBMAd counterparts. Mouse models have been used for BMAds research, but isolation of primary BMAds presents many challenges, and thus in vitro models remain the current standard to study nuances of adipocyte differentiation. No in vitro model has yet been described for the study of rBMAds/cBMAds. Here, we present an in vitro model of BM adipogenesis with differential rBMAd and cBMAd-like characteristics. We used OP9 BM stromal cells derived from a (C57BL/6xC3H)F2-op/op mouse, which have been extensively characterized as feeder layer for hematopoiesis research. We observed similar canonical adipogenesis transcriptional signatures for spontaneously-differentiated (sOP9) and induced (iOP9) cultures, while fatty acid composition and desaturase expression of Scd1 and Fads2 differed at the population level. To resolve differences at the single adipocyte level we tested Raman microspectroscopy and show it constitutes a high-resolution method for studying adipogenesis in vitro in a label-free manner, with resolution to individual LDs. We found sOP9 adipocytes have lower unsaturation ratios, smaller LDs and higher hematopoietic support than iOP9 adipocytes, thus functionally resembling rBMAds, while iOP9 more closely resembled cBMAds. Validation in human primary samples confirmed a higher unsaturation ratio for lipids extracted from stable cBMAd-rich sites (femoral head upon hip-replacement surgery) versus labile rBMAds (iliac crest after chemotherapy). As a result, the 16:1/16:0 fatty acid unsaturation ratio, which was already shown to discriminate BMAd subtypes in rabbit and rat marrow, was validated to discriminate cBMAds from rBMAd in both the OP9 model in vitro system and in human samples. We expect our model will be useful for cBMAd and rBMAd studies, particularly where isolation of primary BMAds is a limiting step. |
| first_indexed | 2024-04-11T15:02:35Z |
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| id | doaj.art-4902e5f201334c0e8c7482fd57750abb |
| institution | Directory Open Access Journal |
| issn | 1664-2392 |
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| last_indexed | 2024-04-11T15:02:35Z |
| publishDate | 2022-11-01 |
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| series | Frontiers in Endocrinology |
| spelling | doaj.art-4902e5f201334c0e8c7482fd57750abb2022-12-22T04:16:55ZengFrontiers Media S.A.Frontiers in Endocrinology1664-23922022-11-011310.3389/fendo.2022.10012101001210Raman microspectroscopy reveals unsaturation heterogeneity at the lipid droplet level and validates an in vitro model of bone marrow adipocyte subtypesJosefine Tratwal0Guillaume Falgayrac1Alexandrine During2Nicolas Bertheaume3Charles Bataclan4Daniel N. Tavakol5Vasco Campos6Ludovic Duponchel7George Q. Daley8Guillaume Penel9Christophe Chauveau10Olaia Naveiras11Olaia Naveiras12Olaia Naveiras13Laboratory of Regenerative Hematopoiesis, Ecole Polytechnique Fédérale de Lausanne (EPFL) & Department of Biomedical Sciences, University of Lausanne (UNIL), Lausanne, SwitzerlandUniv. Lille, CHU Lille, Univ. Littoral Côte d’Opale, ULR 4490 - MABLab- Marrow Adiposity Laboratory, Lille, FranceUniv. Lille, CHU Lille, Univ. Littoral Côte d’Opale, ULR 4490 - MABLab- Marrow Adiposity Laboratory, Lille, FranceUniv. Lille, CHU Lille, Univ. Littoral Côte d’Opale, ULR 4490 - MABLab- Marrow Adiposity Laboratory, Lille, FranceLaboratory of Regenerative Hematopoiesis, Ecole Polytechnique Fédérale de Lausanne (EPFL) & Department of Biomedical Sciences, University of Lausanne (UNIL), Lausanne, SwitzerlandLaboratory of Regenerative Hematopoiesis, Ecole Polytechnique Fédérale de Lausanne (EPFL) & Department of Biomedical Sciences, University of Lausanne (UNIL), Lausanne, SwitzerlandLaboratory of Regenerative Hematopoiesis, Ecole Polytechnique Fédérale de Lausanne (EPFL) & Department of Biomedical Sciences, University of Lausanne (UNIL), Lausanne, SwitzerlandUniv. Lille, CNRS, UMR 8516 - LASIRe - Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l’Environnement, Lille, FranceDivision of Hematology/Oncology, Boston Children’s Hospital and Dana Farber Cancer Institute, Boston, Boston, MA, United StatesUniv. Lille, CHU Lille, Univ. Littoral Côte d’Opale, ULR 4490 - MABLab- Marrow Adiposity Laboratory, Lille, FranceUniv. Lille, CHU Lille, Univ. Littoral Côte d’Opale, ULR 4490 - MABLab- Marrow Adiposity Laboratory, Lille, FranceLaboratory of Regenerative Hematopoiesis, Ecole Polytechnique Fédérale de Lausanne (EPFL) & Department of Biomedical Sciences, University of Lausanne (UNIL), Lausanne, SwitzerlandService of Hematology, Department of Oncology, Lausanne University Hospital and University of Lausanne, Lausanne, SwitzerlandService of Hematology, Department of Laboratory Medicine Oncology, Lausanne University Hospital and University of Lausanne, Lausanne, SwitzerlandBone marrow adipocytes (BMAds) constitute the most abundant stromal component of adult human bone marrow. Two subtypes of BMAds have been described, the more labile regulated adipocytes (rBMAds) and the more stable constitutive adipocytes (cBMAds), which develop earlier in life and are more resilient to environmental and metabolic disruptions. In vivo, rBMAds are enriched in saturated fatty acids, contain smaller lipid droplets (LDs) and more readily provide hematopoietic support than their cBMAd counterparts. Mouse models have been used for BMAds research, but isolation of primary BMAds presents many challenges, and thus in vitro models remain the current standard to study nuances of adipocyte differentiation. No in vitro model has yet been described for the study of rBMAds/cBMAds. Here, we present an in vitro model of BM adipogenesis with differential rBMAd and cBMAd-like characteristics. We used OP9 BM stromal cells derived from a (C57BL/6xC3H)F2-op/op mouse, which have been extensively characterized as feeder layer for hematopoiesis research. We observed similar canonical adipogenesis transcriptional signatures for spontaneously-differentiated (sOP9) and induced (iOP9) cultures, while fatty acid composition and desaturase expression of Scd1 and Fads2 differed at the population level. To resolve differences at the single adipocyte level we tested Raman microspectroscopy and show it constitutes a high-resolution method for studying adipogenesis in vitro in a label-free manner, with resolution to individual LDs. We found sOP9 adipocytes have lower unsaturation ratios, smaller LDs and higher hematopoietic support than iOP9 adipocytes, thus functionally resembling rBMAds, while iOP9 more closely resembled cBMAds. Validation in human primary samples confirmed a higher unsaturation ratio for lipids extracted from stable cBMAd-rich sites (femoral head upon hip-replacement surgery) versus labile rBMAds (iliac crest after chemotherapy). As a result, the 16:1/16:0 fatty acid unsaturation ratio, which was already shown to discriminate BMAd subtypes in rabbit and rat marrow, was validated to discriminate cBMAds from rBMAd in both the OP9 model in vitro system and in human samples. We expect our model will be useful for cBMAd and rBMAd studies, particularly where isolation of primary BMAds is a limiting step.https://www.frontiersin.org/articles/10.3389/fendo.2022.1001210/fullfatty acidunsaturation (fatty acid)OP9 celllipid droplet diameterregulated bone marrow adiposityconstitutive bone marrow adiposity |
| spellingShingle | Josefine Tratwal Guillaume Falgayrac Alexandrine During Nicolas Bertheaume Charles Bataclan Daniel N. Tavakol Vasco Campos Ludovic Duponchel George Q. Daley Guillaume Penel Christophe Chauveau Olaia Naveiras Olaia Naveiras Olaia Naveiras Raman microspectroscopy reveals unsaturation heterogeneity at the lipid droplet level and validates an in vitro model of bone marrow adipocyte subtypes Frontiers in Endocrinology fatty acid unsaturation (fatty acid) OP9 cell lipid droplet diameter regulated bone marrow adiposity constitutive bone marrow adiposity |
| title | Raman microspectroscopy reveals unsaturation heterogeneity at the lipid droplet level and validates an in vitro model of bone marrow adipocyte subtypes |
| title_full | Raman microspectroscopy reveals unsaturation heterogeneity at the lipid droplet level and validates an in vitro model of bone marrow adipocyte subtypes |
| title_fullStr | Raman microspectroscopy reveals unsaturation heterogeneity at the lipid droplet level and validates an in vitro model of bone marrow adipocyte subtypes |
| title_full_unstemmed | Raman microspectroscopy reveals unsaturation heterogeneity at the lipid droplet level and validates an in vitro model of bone marrow adipocyte subtypes |
| title_short | Raman microspectroscopy reveals unsaturation heterogeneity at the lipid droplet level and validates an in vitro model of bone marrow adipocyte subtypes |
| title_sort | raman microspectroscopy reveals unsaturation heterogeneity at the lipid droplet level and validates an in vitro model of bone marrow adipocyte subtypes |
| topic | fatty acid unsaturation (fatty acid) OP9 cell lipid droplet diameter regulated bone marrow adiposity constitutive bone marrow adiposity |
| url | https://www.frontiersin.org/articles/10.3389/fendo.2022.1001210/full |
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