Macrophage Identification In Situ

Nikovics, Krisztina; Favier, Anne-Laure

doi:10.3390/biomedicines9101393

Cited by 14 publications

(17 citation statements)

References 143 publications

(211 reference statements)

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“…These methods only provide a result from a mixture of different cell types. In addition, measuring the expression level of the cytokines is not sufficient; the localization of the cytokine-expressing cells in the tissue should also be determined [ 47 ].…”

Section: Introductionmentioning

confidence: 99%

“…This technique can only distinguish between M1-like and M2-like macrophages. However, it is not appropriate for identifying M2-like subtypes, as there is currently no cell surface marker available to distinguish between the different subtypes; and (iii) in situ hybridization is one of the most convincing methods to identify the cytokine-expressing cells because it is based on messenger RNA (mRNA) detection of the targeted genes [ 47 ].…”

Section: Introductionmentioning

confidence: 99%

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Macrophages Characterization in an Injured Bone Tissue

Nikovics¹,

Durand

Castellarin³

et al. 2022

Biomedicines

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Biomaterial use is a promising approach to facilitate wound healing of the bone tissue. Biomaterials induce the formation of membrane capsules and the recruitment of different types of macrophages. Macrophages are immune cells that produce diverse combinations of cytokines playing an important role in bone healing and regeneration, but the exact mechanism remains to be studied. Our work aimed to identify in vivo macrophages in the Masquelet induced membrane in a rat model. Most of the macrophages in the damaged area were M2-like, with smaller numbers of M1-like macrophages. In addition, high expression of IL-1β and IL-6 cytokines were detected in the membrane region by RT-qPCR. Using an innovative combination of two hybridization techniques (in situ hybridization and in situ hybridization chain reaction (in situ HCR)), M2b-like macrophages were identified for the first time in cryosections of non-decalcified bone. Our work has also demonstrated that microspectroscopical analysis is essential for macrophage characterization, as it allows the discrimination of fluorescence and autofluorescence. Finally, this work has revealed the limitations of immunolabelling and the potential of in situ HCR to provide valuable information for in vivo characterization of macrophages.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Macrophages Characterization in an Injured Bone Tissue

Nikovics¹,

Durand

Castellarin³

et al. 2022

Biomedicines

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“…In the 1980s, the fluorescence in situ hybridization (FISH) technique was published [ 24 ]. This method has the advantage of allowing the simultaneous analysis of several genes in the same sample, but is only suitable for the analysis of highly expressed genes [ 25 ]. Recently, a new method has been published in which in situ hybridization is coupled with the detection by hybridization chain reaction (in situ-HCR) [ 21 , 26 , 27 , 28 ].…”

Section: Introductionmentioning

confidence: 99%

“…In addition, several genes can be analyzed simultaneously. However, in situ-HCR is less efficient than the in situ-DIG method [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

In Situ Gene Expression in Native Cryofixed Bone Tissue

Nikovics¹,

Castellarin²,

Holy³

et al. 2022

Biomedicines

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Bone is a very complex tissue that is constantly changing throughout the lifespan. The precise mechanism of bone regeneration remains poorly understood. Large bone defects can be caused by gunshot injury, trauma, accidents, congenital anomalies and tissue resection due to cancer. Therefore, understanding bone homeostasis and regeneration has considerable clinical and scientific importance in the development of bone therapy. Macrophages are well known innate immune cells secreting different combinations of cytokines and their role in bone regeneration during bone healing is essential. Here, we present a method to identify mRNA transcripts in cryosections of non-decalcified rat bone using in situ hybridization and hybridization chain reaction to explore gene expression in situ for better understanding the gene expression of the bone tissues.

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“…Current methods related to the identification of macrophages in situ were systematized and discussed, including in situ hybridization, immunolabeling and other approaches [ 22 ].…”

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confidence: 99%