Efficient primary culture model of patient‑derived tumor cells from colorectal cancer using a Rho‑associated protein kinase inhibitor and feeder cells

Hong, Hye Kyung; Pyo, Dae Hee; Kim, Tae Won; Yun, Nak Hyeon; Lee, Yeo Song; Song, Su Jeong; Lee, Woo Yong; Cho, Yong Beom

doi:10.3892/or.2019.7285

Cited by 5 publications

(4 citation statements)

References 41 publications

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“…There has been literature that reported characterization of spheroids established from lung cancer cell lines or xenografts [ 23 , 45 ]. The list of the genes or pathways that change during the formation of spheroids were partially similar but partially different from ours.…”

Section: Discussionmentioning

confidence: 99%

Patient-derived spheroids and patient-derived organoids simulate evolutions of lung cancer

Surina

Tanggis

Suzuki

et al. 2023

Heliyon

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Section: Discussionmentioning

confidence: 99%

Patient-derived spheroids and patient-derived organoids simulate evolutions of lung cancer

Surina

Tanggis

Suzuki

et al. 2023

Heliyon

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“…However, an efficient in vitro primary culture system for hHFSCs is currently not established. The main factors affecting the proliferation and biological characteristics of an adherent culture in vitro are the coated culture dishes and culture medium (Hong et al, 2019;Hashimoto et al, 2020). In this study, we obtained hHFSCs using a two-step proteolytic digestion combined with microscopy.…”

Section: Discussionmentioning

confidence: 99%

Establishment of an Efficient Primary Culture System for Human Hair Follicle Stem Cells Using the Rho-Associated Protein Kinase Inhibitor Y-27632

Wen

Miao

Fan

et al. 2021

Front. Cell Dev. Biol.

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BackgroundHair follicle tissue engineering is a promising strategy for treating hair loss. Human hair follicle stem cells (hHFSCs), which play a key role in the hair cycle, have potential applications in regenerative medicine. However, previous studies did not achieve efficient hHFSC expansion in vitro using feeder cells. Therefore, there is a need to develop an efficient primary culture system for the expansion and maintenance of hHFSCs.MethodsThe hHFSCs were obtained by two-step proteolytic digestion combined with microscopy. The cell culture dishes were coated with human fibronectin and inoculated with hHFSCs. The hHFSCs were harvested using a differential enrichment procedure. The effect of Rho-associated protein kinase (ROCK) inhibitor Y-27632, supplemented in keratinocyte serum-free medium (K-SFM), on adhesion, proliferation, and stemness of hHFSCs and the underlying molecular mechanisms were evaluated.ResultsThe hHFSCs cultured in K-SFM, supplemented with Y-27632, exhibited enhanced adhesion and proliferation. Additionally, Y-27632 treatment maintained the stemness of hHFSCs and promoted the ability of hHFSCs to regenerate hair follicles in vivo. However, Y-27632-induced proliferation and stemness in hHFSCs were conditional and reversible. Furthermore, Y-27632 maintained propagation and stemness of hHFSCs through the ERK/MAPK pathway.ConclusionAn efficient short-term culture system for primary hHFSCs was successfully established using human fibronectin and the ROCK inhibitor Y-27632, which promoted the proliferation, maintained the stemness of hHFSCs and promoted the ability to regenerate hair follicles in vivo. The xenofree culturing method used in this study provided a large number of high-quality seed cells, which have applications in hair follicle tissue engineering and stem cell therapy.

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“…Patient-derived cancer cells (PDCs) were established, and the genetic identification and culture method were described previously. 14 15 Bone marrow-derived macrophages or dendritic cells (BMDMs or BMDCs) differentiation method and cell viability and apoptosis analysis method were described in detail in the online supplemental material .…”

Section: Methodsmentioning

confidence: 99%

Statin-mediated inhibition of RAS prenylation activates ER stress to enhance the immunogenicity of KRAS mutant cancer

Nam

Kwon

Jung

et al. 2021

J Immunother Cancer

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BackgroundStatins preferentially promote tumor-specific apoptosis by depleting isoprenoid such as farnesyl pyrophosphate and geranylgeranyl pyrophosphate. However, statins have not yet been approved for clinical cancer treatment due, in part, to poor understanding of molecular determinants on statin sensitivity. Here, we investigated the potential of statins to elicit enhanced immunogenicity of KRAS-mutant (KRASmut) tumors.MethodsThe immunogenicity of treated cancer cells was determined by western blot, flow cytometry and confocal microscopy. The immunotherapeutic efficacy of mono or combination therapy using statin was assessed in KRASmut tumor models, including syngeneic colorectal cancer and genetically engineered lung and pancreatic tumors. Using NanoString analysis, we analyzed how statin influenced the gene signatures associated with the antigen presentation of dendritic cells in vivo and evaluated whether statin could induce CD8+ T-cell immunity. Multiplex immunohistochemistry was performed to better understand the complicated tumor-immune microenvironment.ResultsStatin-mediated inhibition of KRAS prenylation provoked severe endoplasmic reticulum (ER) stress by attenuating the anti-ER stress effect of KRAS mutation, thereby resulting in the immunogenic cell death (ICD) of KRASmut cancer cells. Moreover, statin-mediated ICD enhanced the cross-priming ability of dendritic cells, thereby provoking CD8+ T-cell immune responses against KRASmut tumors. Combination therapy using statin and oxaliplatin, an ICD inducer, significantly enhanced the immunogenicity of KRASmut tumors and promoted tumor-specific immunity in syngeneic and genetically engineered KRASmut tumor models. Along with immune-checkpoint inhibitors, the abovementioned combination therapy overcame resistance to PD-1 blockade therapies, improving the survival rate of KRASmut tumor models.ConclusionsOur findings suggest that KRAS mutation could be a molecular target for statins to elicit potent tumor-specific immunity.

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Efficient primary culture model of patient‑derived tumor cells from colorectal cancer using a Rho‑associated protein kinase inhibitor and feeder cells

Cited by 5 publications

References 41 publications

Patient-derived spheroids and patient-derived organoids simulate evolutions of lung cancer

Patient-derived spheroids and patient-derived organoids simulate evolutions of lung cancer

Establishment of an Efficient Primary Culture System for Human Hair Follicle Stem Cells Using the Rho-Associated Protein Kinase Inhibitor Y-27632

Statin-mediated inhibition of RAS prenylation activates ER stress to enhance the immunogenicity of KRAS mutant cancer

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