Evaluation of phenotypic and functional stability of RAW 264.7 cell line through serial passages

Taciak, Bartłomiej; Białasek, Maciej; Braniewska, Agata; Sas, Zuzanna; Sawicka, Paulina; Kiraga, Łukasz; Rygiel, Tomasz P.; Król, Magdalena

doi:10.1371/journal.pone.0198943

Cited by 216 publications

(176 citation statements)

References 28 publications

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“…Therefore, combined with previous excellent and novel studies in these fields, 4,32 in our experiences, it might be a reason for different RAW-OC induction efficacies among various study groups. 32 Interestingly, however, as an iNOS inhibitory genes, the Arg1 also highly expressed in another polarized macrophage stage: anti-inflammatory (M2) phenotype, 49,50 and its expression in RAW264.7 has be showed significantly higher in the passage of No. 32 Collin-Osdoby et al 4 32 However, comparing to RAW264.7, the bone marrow macrophage-M0 (BMM-Mφ) has been reported markedly express CD169.…”

Section: Phenot Ype S Tudy Of R Aw26 47 and Its Role On The Reg Ulmentioning

confidence: 99%

“…On the other hand, their stability between various laboratories and passages is questionable. 4,19,32 Otherwise, previous study showed the elder RAW264.7 cells could change its morphology and decrease the production of proteins, which lead RAW264.7 cells resistant for differentiation and transduction. 18, since study showing the induction efficacy might decrease with passage and consequently phenotype changing.…”

Section: Phenot Ype S Tudy Of R Aw26 47 and Its Role On The Reg Ulmentioning

confidence: 99%

“…18, since study showing the induction efficacy might decrease with passage and consequently phenotype changing. 32 Collin-Osdoby et al 4 32 However, comparing to RAW264.7, the bone marrow macrophage-M0 (BMM-Mφ) has been reported markedly express CD169. 44 However, there still lack evidence for rigid classification criteria of cell lines between various passages and it seems to be cell-linage dependent.…”

Section: Phenot Ype S Tudy Of R Aw26 47 and Its Role On The Reg Ulmentioning

confidence: 99%

“…32 Collin-Osdoby et al 4 32 However, comparing to RAW264.7, the bone marrow macrophage-M0 (BMM-Mφ) has been reported markedly express CD169. 32 Notably, the expression of TRAP, which involved in osteoclastogenesis and as one of osteoclastic specific genes remains stable in RAW264.7 cell linage from the passage No. 48 Similarly, genes in subpanel-2 and subpanel-3 also associated with macrophage activation including: CD86, HIF-1α, CD11a, CD18, CD206, CD200R, Glut1 (Glucose transporter 1) and Ly6c and TfR2, Arg1 and SCARA-5b (scavenger receptor class A member 5) respectively.…”

Section: Phenot Ype S Tudy Of R Aw26 47 and Its Role On The Reg Ulmentioning

confidence: 99%

“…[45][46][47] Otherwise, the CD11c and iNOS in subpanel-1 are demonstrated as highly expressed in pro-inflammatory macrophage stage (M1). 32 Besides that, Wang et al 51 have reported that CD109, as a glycosyl-phosphatidylinositol (GPI)-anchored protein play a crucial role in RANKL-induced RAW-OCs formation and bone resorptive function. 32 Interestingly, however, as an iNOS inhibitory genes, the Arg1 also highly expressed in another polarized macrophage stage: anti-inflammatory (M2) phenotype, 49,50 and its expression in RAW264.7 has be showed significantly higher in the passage of No.…”

Section: Phenot Ype S Tudy Of R Aw26 47 and Its Role On The Reg Ulmentioning

confidence: 99%

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Overview of RAW264.7 for osteoclastogensis study: Phenotype and stimuli

Kong

Smith

Hao

2019

J Cellular Molecular Medi

109

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Bone homeostasis is preserved by the balance of maintaining between the activity of osteogenesis and osteoclastogenesis. However, investigations for the osteoclastogenesis were hampered by considerable difficulties associated with isolating and culturing osteoclast in vivo. As the alternative, stimuli‐induced osteoclasts formation from RAW264.7 cells (RAW‐OCs) have gain its importance for extensively osteoclastogenic study of bone diseases, such as rheumatoid arthritis, osteoporosis, osteolysis and periodontitis. However, considering the RAW‐OCs have not yet been well‐characterized and RAW264.7 cells are polymorphic because of a diverse phenotype of the individual cells comprising this cell linage, and different fate associated with various stimuli contributions. Thus, in present study, we provide an overview for current knowledge of the phenotype of RAW264.7 cells, as well as the current understanding of the complicated interactions between various stimuli and RAW‐OCs in the light of the recent progress.

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Section: Phenot Ype S Tudy Of R Aw26 47 and Its Role On The Reg Ulmentioning

confidence: 99%

Section: Phenot Ype S Tudy Of R Aw26 47 and Its Role On The Reg Ulmentioning

confidence: 99%

Section: Phenot Ype S Tudy Of R Aw26 47 and Its Role On The Reg Ulmentioning

confidence: 99%

Section: Phenot Ype S Tudy Of R Aw26 47 and Its Role On The Reg Ulmentioning

confidence: 99%

Section: Phenot Ype S Tudy Of R Aw26 47 and Its Role On The Reg Ulmentioning

confidence: 99%

See 3 more Smart Citations

Overview of RAW264.7 for osteoclastogensis study: Phenotype and stimuli

Kong

Smith

Hao

2019

J Cellular Molecular Medi

109

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MyD88 and not TRIF knockout is sufficient to abolish LPS‐induced inflammatory responses in bone‐derived macrophages

et al. 2023

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Macrophages play an important role in the response to infection and/or repair of injury in tissues. To examine the NF‐κB pathway in response to an inflammatory stimulus, we used wild‐type bone‐marrow‐derived macrophages (BMDMs) or BMDMs with knockout (KO) of myeloid differentiation primary response 88 (MyD88) and/or Toll/interleukin‐1 receptor domain‐containing adapter‐inducing interferon‐β (TRIF) via CRISPR/Cas9. Following treatment of BMDMs with lipopolysaccharide (LPS) to induce an inflammatory response, translational signalling of NF‐κB was quantified via immunoblot and cytokines were measured. Our findings reveal that MyD88 KO, but not TRIF KO, decreased LPS‐induced NF‐κB signalling, and 10% expression of basal MyD88 expression was sufficient to partially rescue the abolished inflammatory cytokine secretion observed upon MyD88 KO.

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Multitargeted Nanoparticles Deliver Synergistic Drugs across the Blood–Brain Barrier to Brain Metastases of Triple Negative Breast Cancer Cells and Tumor‐Associated Macrophages

Zhang

Lip

et al. 2019

Adv Healthcare Materials

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Owing to the low expression of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 (HER2), endocrine and anti-HER2 therapies are ineffective against TNBC. Thus currently, standard adjuvant and neoadjuvant treatments of TNBC are limited to conventional anthracycline-taxanebased chemotherapy. [1] Despite the initial response to chemotherapy, TNBC patients have a high risk of relapse and distal metastases, especially to the brain, leading to shortened survival times. [2] A number of targeted therapies have been investigated to treat TNBC including poly(ADPribose) polymerase (PARP) inhibitors, immune checkpoint inhibitors, antiandrogen agents, phosphoinositide 3-kinase inhibitors, mitogen-activated protein kinase inhibitors, antiangiogenic antibody, and integrin inhibitors. [1,3] Upregulation of cell surface αvβ3 and αvβ5 integrins in aggressive TNBC, especially in brain metastases and tumorassociated vasculature, has presented an opportunity for targeted therapy of TNBC using integrin-targeted peptides. [4] Of various peptide-based inhibitors of αv integrins, Cilengitide, an Arg-Gly-Asp (RGD) peptide mimetic, with high selectivity against integrins αvβ3 and αvβ5, reached phase 3 clinical trial; however, the results were negative. [5] This disappointing outcome may be caused by insufficient amounts of inhibitor reaching tumor cells to generate a direct cytotoxic effect against αvβ3 and αvβ5 positive cancer cells due to the short half-life of Cilengitide in vivo. [5] Nonetheless, RGD-conjugated nanoparticulate drug delivery systems have shown an ability to target chemotherapeutic drugs to integrin-overexpressing tumor vasculature and cancer cells. [6] Despite the promising results in preclinical models, it is challenging to find an optimal RGD particle coverage to balance tumor accumulation against liver uptake. For example, our previous studies on cyclic RGDfK (Arg-Gly-Asp-D-Phe-Lys; cRGD) peptide-conjugated solid-lipid nanoparticles (SLN) or polymer-lipid hybrid nanoparticles (PLN) revealed that a high density of cRGD on the surface led to increased liver uptake Patients with brain metastases of triple negative breast cancer (TNBC) have a poor prognosis owing to the lack of targeted therapies, the aggressive nature of TNBC, and the presence of the blood-brain barrier (BBB) that blocks penetration of most drugs. Additionally, infiltration of tumor-associated macrophages (TAMs) promotes tumor progression. Here, a terpolymer-lipid hybrid nanoparticle (TPLN) system is designed with multiple targeting moieties to first undergo synchronized BBB crossing and then actively target TNBC cells and TAMs in microlesions of brain metastases. In vitro and in vivo studies demonstrate that covalently bound polysorbate 80 in the terpolymer enables the low-density lipoprotein receptor-mediated BBB crossing and TAM-targetability of the TPLN. Conjugation of cyclic internalizing peptide (iRGD) enhances cellular uptake, cytotoxicity, and drug delivery to brain metastases of integrinoverexpressing TNB...

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Evaluation of phenotypic and functional stability of RAW 264.7 cell line through serial passages

Cited by 216 publications

References 28 publications

Overview of RAW264.7 for osteoclastogensis study: Phenotype and stimuli

Overview of RAW264.7 for osteoclastogensis study: Phenotype and stimuli

MyD88 and not TRIF knockout is sufficient to abolish LPS‐induced inflammatory responses in bone‐derived macrophages

Multitargeted Nanoparticles Deliver Synergistic Drugs across the Blood–Brain Barrier to Brain Metastases of Triple Negative Breast Cancer Cells and Tumor‐Associated Macrophages

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