Establishment and characterization of two novel patient-derived cell lines from giant cell tumor of bone: NCC-GCTB8-C1 and NCC-GCTB9-C1

doi:10.21203/rs.3.rs-3457371/v1

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Establishment and characterization of two novel patient-derived cell lines from giant cell tumor of bone: NCC-GCTB8-C1 and NCC-GCTB9-C1

https://doi.org/10.21203/rs.3.rs-3457371/v1

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published 11 Mar, 2024

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Giant cell tumor of bone (GCTB) is a rare osteolytic bone tumor consisting of mononuclear stromal cells, macrophages, and osteoclast-like giant cells. Although GCTB predominantly exhibits benign behavior, the tumor carries a significant risk of high local recurrence. Furthermore, GCTB can occasionally undergo malignant transformation and distal metastasis, making it potentially fatal. The standard treatment is complete surgical resection; nonetheless, an optimal treatment strategy for advanced GCTB remains unestablished, necessitating expanded preclinical research to identify appropriate therapeutic options. However, only one GCTB cell line is publicly available from a cell bank for research use worldwide. The present study reports the establishment of two novel cell lines, NCC-GCTB8-C1 and NCC-GCTB9-C1, derived from the primary tumor tissues of two patients with GCTB. Both cell lines maintained the hallmark mutation in the H3-3A gene, which is associated with tumor formation and development in GCTB. Characterization of these cell lines revealed their steady growth, spheroid-formation capability, and invasive traits. Potential therapeutic agents were identified via extensive drug screening of the two cell lines and seven previously established GCTB cell lines. Among the 214 antitumor agents tested, romidepsin, a histone deacetylase inhibitor, and mitoxantrone, a topoisomerase inhibitor, were identified as potential therapeutic agents against GCTB. Conclusively, the establishment of NCC-GCTB8-C1 and NCC-GCTB9-C1 provides novel and crucial resources that are expected to advance GCTB research and potentially revolutionize treatment strategies.

Giant cell tumor of bone

Bone tumor

Cell lines

Patient-derived model

Antitumor drug screening

Giant cell tumor of bone (GCTB) is a rare bone tumor composed of neoplastic mononuclear stromal cells with a monotonous appearance mixed with macrophages and osteoclast-like giant cells [1]. GCTB comprises approximately 5% of all primary bone tumors [1, 2]. The estimated incidence of GCTB is 1.2 to 1.7 per million individuals per year and is slightly more common in women [3, 4]. The tumor predominantly appears between the ages of 20 and 45 years; however, approximately 10% of these cases occur during the second decade of life [5]. GCTB often occurs in the epiphysis of long bones, including the distal femur, proximal tibia, distal radius, and proximal humerus. Metaphysis can also arise in some cases [1]. Genetically, mutations in the H3-3A G34W gene are found in nearly 90% of GCTBs, suggesting its crucial role in tumor formation. Several reports have indicated the relationship between this mutant histone and the pathogenesis of GCTB; nevertheless, its precise function remains unclear [1, 6–8].

Symptoms such as pain, swelling, potential joint movement restrictions, and pathologic fractures are reported in 5–12% of patients with GCTB [1]. Imaging studies typically display lytic and expansile lesions, although a definitive diagnosis is usually based on pathological examination of surgery or biopsy specimens [3, 5, 9]. GCTB is primarily treated via surgery, which aims to achieve maximum tumor removal and curettage with bone preservation. However, for unresectable tumors or surgeries with a risk of high morbidity, denosumab, a receptor activator of nuclear factor kappa-B ligand inhibitor, is a reasonable therapeutic option [2, 4, 10]. Although the 5-year survival rate is approximately 90% and the general prognosis is favorable, GCTB has several clinical problems [11, 12]. First, the local recurrence rate is high (15–50%) two years postoperatively [1, 4, 13]; second, malignant transformation (1.4–6.6%) and pulmonary metastases (1–6%) can be fatal [13–15]; third, the optimal administration method of denosumab for advanced GCTB cases has not yet been established [5, 10]. Therefore, developing new therapeutic agents for GCTB is pertinent.

Patient-derived cell lines are essential research resources for rare tumors because enrolling enough patients in clinical trials can seldom be accomplished within a reasonable time [16]. However, the number of bone tumor cell lines is insufficient compared to that in major cancers, and even fewer in GCTB [17, 18]. Based on the Cellosaurus cell line database, 13 GCTB cell lines have been reported (Supplementary Table 1) [19]. Nevertheless, researchers have access to only one of them, which has been submitted to cell banks currently. Therefore, additional GCTB cell lines are crucial for GCTB research. In this context, the present study aimed at establishing GCTB cell lines from tumor tissues obtained from patients with GCTB. The findings of this study are expected to favor the development of novel therapeutic strategies for GCTB.

2.1 Patient data

Case 1

The donor patient was a 32-year-old man with a GCTB. The patient had no relevant medical history. He experienced pain in his left knee while exercising, but it was untreated for a year. The pain gradually worsened, prompting the patient to visit a hospital. Imaging studies revealed findings suggestive of a bone tumor in the proximal tibia. He was then referred to the National Cancer Center Hospital in Tokyo, Japan, for further evaluation and treatment. Radiography and computed tomography (CT) scans showed an osteolytic lesion with marginal sclerosis in the proximal tibia (Fig. 1A, B). Magnetic resonance imaging (MRI) confirmed the presence of a neoplastic lesion in the proximal tibia, showing heterogeneous low signal intensity on the T2-weighted image (Fig. 1C) and heterogeneous high signal intensity on the gadolinium-enhanced T1-weighted image (Fig. 1D). The tumor was diagnosed as a GCTB based on open biopsy, and no distal or lymph node metastases were detected. The patient underwent aggressive curettage of the lesion, with reinforcement using autologous bone graft implantation and plate fixation. Tumor tissues obtained during the surgery were used to establish cell lines. Histologically, the tumor consisted of numerous osteoclast-like giant cells and mononuclear non-atypical tumor cells with hemosiderin-laden macrophages and hemorrhages (Fig. 1E). The neoplastic cells showed nuclear immunohistochemical expression of H3-3A G34W (Fig. 1F). The patient experienced an uneventful postoperative course, and neither local recurrence nor metastasis were observed. The use of clinical materials for this study was approved by the ethics committee of the National Cancer Center, and written informed consent was obtained from the donor patient.

Case 2

The donor patient was a 33-year-old woman who was diagnosed with GCTB. The patient had no relevant medical history. She presented with right ankle pain and sought medical attention. Imaging revealed findings suggestive of a bone tumor in the distal tibia. Subsequently, she was referred to the National Cancer Center Hospital in Tokyo, Japan, for further evaluation and treatment. Radiography and CT revealed an osteolytic lesion with destructive changes in the distal tibia (Fig. 2A, B). MRI confirmed the presence of a neoplastic lesion in the distal tibia, displaying heterogeneous low-signal intensity on the T2-weighted image (Fig. 2C) and iso-signal intensity on the T2-weighted fat saturation image (Fig. 2D). The tumor was diagnosed as a GCTB based on a needle biopsy under imaging guidance. No distant or lymph node metastases were observed. The patient underwent aggressive curettage of the lesion with reinforcement using autologous and artificial bone graft implantation and plate fixation. Histologically, the tumor consisted of uniform round-to-oval neoplastic cells and osteoclast-like multinucleated giant cells (Fig. 2E). Immunohistochemical examination revealed that the mononuclear cells were diffusely positive for H3-3A G34W (Fig. 2F). The patient experienced an uneventful postoperative course, and neither local recurrence nor metastasis were observed. The use of clinical materials for this study was approved by the ethics committee of the National Cancer Center, and written informed consent was obtained from the donor patient.

2.2 Histological analysis

A histological assessment was conducted on 4 µm-thick sections derived from a typical tumor sample and embedded in paraffin. The slices were deparaffinized and stained with hematoxylin and eosin (H & E). Further staining with the H3-3A G34W antibody (1:1000, RM263; RevMAb Biosciences USA, Inc., CA, USA) was performed to investigate the expression of H3-3A G34W.

2.3 Establishment of cell lines

Primary GCTB tumor cells were obtained from surgically removed tumor tissues and subsequently cut into small fragments using scissors. The gentleMACS™ Octo Dissociator with Heaters (Miltenyi Biotec B.V. & Co. KG, Bergisch Gladbach, Germany) was used to digest and homogenize the dissected tissues within the gentleMACS™ C Tube (Miltenyi Biotec) for 30 min at 37°C using the Tumor Cell Dissociation Kit, human (Miltenyi Biotec). The derived cells were then placed on 60 mm collagen type I-coated culture plates (Sumitomo Bakelite Co. Ltd., Tokyo, Japan). Sustenance of the cells was achieved through DMEM/F-12 medium (Thermo Fisher Scientific Inc., MA, USA) enriched with GlutaMAX (Thermo Fisher Scientific Inc.), 5% heat-inactivated fetal bovine serum (FBS) (Thermo Fisher Scientific Inc.), and other specified compounds. The culture medium was replaced every two–three days. A microscope (Carl Zeiss AG, Land Baden-Württemberg, Germany) was used to monitor cell confluence. Once subconfluency was attained, the cultured cells were washed with phosphate-buffered saline (–) (Nacalai Tesque Inc., Kyoto, Japan) and subsequently dissociated with trypsin–EDTA solution (Nacalai Tesque Inc.). The cells were placed in tissue culture plates and maintained at 37°C in an atmosphere containing 5% carbon dioxide.

2.4 Authentication and quality control of the cell lines

For authentication and quality assurance of the developed cell lines, DNA was extracted from the tumor tissues and cell lines. The Wizard® Genomic DNA Purification Kit (Promega Co., WI, USA) and the Qiagen DNeasy Blood and Tissue Kit (QIAGEN N.V., Hilden, Germany) were used for DNA extraction. The DNA concentration was determined using the NanoDrop 8000 (Thermo Fisher Scientific Inc.). The established cell lines were verified via short tandem repeat (STR) analysis across 10 loci using the GenePrint 10 system (Promega Co.) and a 3500xL Genetic Analyzer (Thermo Fisher Scientific Inc.).

2.5 Genetic analysis

Using the QIAzol Lysis Reagent (QIAGEN) and miRNeasy Mini Kit (QIAGEN), GCTB cells were subjected to total RNA extraction to identify gene mutations. The extracted RNA was then converted to complementary DNA using SuperScript III reverse transcriptase (Invitrogen, MA, USA) as per manufacturer’s instructions. The H3-3A gene was amplified via polymerase chain reaction (PCR) using the H3-3A forward primer H3-3A_F (5′-TAA AGCACCCAGGAAGCAAC-3′), H3-3A reverse primer H3-3A_R (5′-CAAGAGAGACTTTGTCCCATTTTT-3′), and Platinum Taq DNA Polymerase High Fidelity (Life Technologies Co., CA, USA). PCR products were purified using the Wizard SV Gel and PCR Clean-Up System (Promega Co.), followed by direct sequencing using the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, MA, USA) and Applied Biosystems 3130xL Genetic Analyzer (Thermo Fisher Scientific Inc.) as conducted by GENEWIZ. Sequence results were analyzed using A Plasmid Editor v2.0.61.

2.6 Inspection of Mycoplasma contamination

For the assessment of Mycoplasma contamination, cellular DNA was analyzed using the conventional method of cell line establishment as defined by the international cell line authentication committee (The International Cell Line Authentication Committee 2018) [20]. Cellular DNA was amplified using an e-Myco Mycoplasma PCR Detection Kit (Intron Biotechnology, Gyeonggi-do, Korea). Thereafter, the DNA was separated on a 1.5% agarose gel via electrophoresis, marked with SYBR Safe DNA gel stain (Invitrogen), and subsequently assessed using Amersham Imager 600 (GE Healthcare Biosciences, Uppsala, Sweden).

2.7 Cell proliferation assay

The cell proliferation ability was assessed using Cell Counting Kit-8 (DOJINDO LABORATORIES, Kumamoto, Japan) every 24 h for four days. The cells were seeded at a density of 1.25 × 10⁴ cells/well into DMEM/F12 enriched with GlutaMAX, 5% FBS, 10 µM Y-27632, 10 ng/mL basic fibroblast growth factor, 5 ng/mL epidermal growth factor, 5 µg/mL insulin, 0.4 µg/mL hydrocortisone, 100 µg/mL penicillin, and 100 µg/mL streptomycin. Doubling time was calculated based on the growth curve.

2.8 Spheroid formation assay

Spheroid formation was evaluated as described previously [21]. The established cells were seeded at a density of 1.0 × 10⁵ cells/well into a 96-well Clear Round Bottom Ultra-Low Attachment Microplate (Corning Inc., NY, USA), and spheroid formation was confirmed via microscopic examination (KEYENCE Co., Osaka, Japan). After a three-day cultivation period, spheroids were retrieved from the plate, encapsulated in a gel using iPGell (GenoStaff Co. Ltd., Tokyo, Japan) and fixed with a 10% solution of neutral formalin buffer. To create paraffin sections of the crafted spheroids, gel-covered formations were enveloped in paraffin and segmented into 4 µm-thick sections. These divided spheroids were then subjected to H & E staining, and subsequently, microscopic inspection was performed.

2.9 Invasion assay

The invasive capabilities of the GCTB cell lines NCC-GCTB8-C1 and NCC-GCTB9-C1 were measured using the real-time cell analyzer xCELLigence (Agilent Technologies Inc., CA, USA). A mixture of DMEM/F12 supplemented with GlutaMAX, 5% FBS, 10 µM Y-27632, 10 ng/mL basic fibroblast growth factor, 5 ng/mL epidermal growth factor, 5 µg/mL insulin, 0.4 µg/mL hydrocortisone, 100 µg/mL penicillin, and 100 µg/mL streptomycin was introduced to the lower chamber. The Matrigel basement membrane matrix (Corning Inc.) was used to coat the membrane in the upper chamber, along with 4.0 × 10⁴ cells suspended in DMEM/F12 devoid of supplements. Cell movement from the upper to lower chamber occurred via the membrane, attaching to the electronic sensors on its underside. The electrical impedance influencing the electronic sensors was monitored using a real-time cell analyzer, with readings taken every 15 min over 72 h. The MG-63 osteosarcoma cell line (Japanese Collection of Research Bioresources Cell Bank, Osaka, Japan) was used as control [22].

2.10 Drug screening assay

The antitumor properties of 214 compounds were evaluated (Supplementary Table 2). Cells were dispersed at a density of 5.0 × 10³ cells/well in 384-well plates and incubated for a day. On the second day, each agent was spread into two wells of the plate and incubated for three days. A Cell Counting Kit-8 (DOUJINDO LABORATORIES) was used to calculate cell viability. The acquired data were consistent with the findings of previous studies on NCC-GCTB1-C1 [6], NCC-GCTB2-C1 [23], NCC-GCTB3-C1 [23], NCC-GCTB4-C1 [21], NCC-GCTB5-C1 [24], NCC-GCTB6-C1 [22], and NCC-GCTB7-C1 [22]. Quantile normalization was performed using R (version 4.0.3, limma package version 3.46.0, Bioconductor), and unsupervised hierarchical clustering was performed using the gplots package (version 3.1.0, CRAN, https://cran.r-project.org).

To determine the half-maximal inhibitory concentration (IC₅₀), 24 compounds exhibiting superior antiproliferative activity based on the initial drug screening test were selected. The cells were treated with each compound at ten distinct concentrations. The IC₅₀ value was derived from logistic regression analysis of the relationship between the common logarithm of the concentration and cell viability for each compound.

3.1 Authentication of the established GCTB cell lines

Cell lines from the patients in Case 1 and Case 2 were successfully established and named NCC-GCTB8-C1 and NCC-GCTB9-C1, respectively. These cell lines were maintained for more than 25 passages over a period of four months. To verify their authenticity, 10 microsatellite loci were analyzed. The STR profiles of the established cell lines matched 100% with those of the original tumor tissues (Table 1, Supplementary Fig. 1). A public database search confirmed that the STR patterns of NCC-GCTB8-C1 and NCC-GCTB9-C1 cells were distinct from those of existing cell lines. These findings support the novelty of the NCC-GCTB8-C1 and NCC-GCTB9-C1 cell lines. Additionally, Mycoplasma contamination test results for these cell lines were negative, as Mycoplasma-specific DNA was not detected (data not shown).

Table 1

Short tandem repeat analysis
Microsatellite	NCC-GCTB8-C1		NCC-GCTB9-C1
(Chromosome)	Cell line	Tumor Tissue (Case 1)	Cell line	Tumor Tissue (Case 2)
Amelogenin (X Y)	X, Y	X, Y	X	X
TH01 (3)	8, 9	8, 9	7, 8	7, 8
D21S11 (21)	31, 32.2	31, 32.2	32	32
D5S818 (5)	12	12	9, 13	9, 13
D13S317 (13)	11, 12	11, 12	12	12
D7S820 (7)	11	11	8, 11	8, 11
D16S539 (16)	9, 10	9, 10	9, 12	9, 12
CSF1PO (5)	10, 12	10, 12	11	11
vWA (12)	14, 17	13, 17	17, 19	17, 19
TPOX (2)	8, 11	8, 11	8	8

3.2 Characterization of the established GCTB cell lines

The presence of the typical GCTB mutation, H3-3A G34W, was detected via Sanger sequencing of NCC-GCTB8-C1 (Fig. 3A) and NCC-GCTB9-C1 cells (Fig. 3B). Both the cell lines displayed adherent characteristics with a spindle-like appearance (Fig. 4A, B). Furthermore, they formed spheroids when cultured on low-attachment plates, with H & E-stained spheroid sections revealing spindle and round cell morphologies but no giant cells (Fig. 4C, D). Growth curve analysis indicated that the NCC-GCTB8-C1 and NCC-GCTB9-C1 cells had population doubling times of 26 h and 12 h, respectively (Fig. 4E). Moreover, both NCC-GCTB8-C1 and NCC-GCTB9-C1 cells demonstrated higher invasive capabilities than the MG63 osteosarcoma cell line (Fig. 4F).

3.3 Sensitivity of the antitumor agents to the established GCTB cell lines

A comprehensive drug assessment was conducted to evaluate the effects of the 214 antitumor agents on the growth of NCC-GCTB8-C1 and NCC-GCTB9-C1 cells. The viability of these cell lines following exposure to each antitumor agent was calculated and is presented in Supplementary Table 3 along with data from seven other GCTB cell lines previously established in our research laboratory. Based on their antitumor effects, the agents were classified into two distinct clusters: cluster A, representing the effective group, and cluster B, representing the ineffective group (Fig. 5A, Supplementary Table 3). Cluster A demonstrated a slightly higher percentage of cytotoxic agents, particularly topoisomerase inhibitors, compared to cluster B (Fig. 5B, C, Supplementary Table 3, 4). The proportion of tyrosine kinases was higher in cluster A than that in cluster B; however, the drug subtypes were equally distributed between clusters A and B (Fig. 5D). Molecularly targeted drugs were similarly represented in clusters A and B, whereas all histone deacetylase (HDAC) inhibitors belonged to cluster A (Fig. 5E).

To further characterize the antitumor agents identified in the screening study, IC₅₀ values were calculated for each agent. The results are presented in Supplementary Table 5, along with the IC₅₀ values for the seven previously established GCTB cell lines. HDAC and topoisomerase inhibitors exhibited remarkable antitumor effects in GCTB cell lines. Among the HDAC inhibitors, romidepsin displayed the most potent antitumor effect, whereas among the topoisomerase inhibitors, mitoxantrone demonstrated the most potent response. The detailed IC₅₀ values and growth curves for these agents are shown in Table 2 and Fig. 6, respectively.

In this study, we successfully established and described two novel cell lines, NCC-GCTB8-C1 and NCC-GCTB9-C1, which were derived from patients with GCTB. Patient-derived cell lines retain genetic and phenotypic characteristics of the original tumor. They serve as invaluable tools for understanding tumor biology, testing drug sensitivities, and modeling potential therapeutic approaches [25–27]. Cell lines are a particularly essential research resource because clinical trials cannot be conducted for tumors with limited cases [16]. However, few GCTB cell lines have been established and only one is currently available for research [19]. Representing the diversity of GCTB clinical behaviors in these cell lines is challenging [2, 4, 28]. Therefore, the establishment of NCC-GCTB8-C1 and NCC-GCTB9-C1 cell lines represents a crucial advancement in broadening the range of available GCTB cell lines for comprehensive research.

NCC-GCTB8-C1 and NCC-GCTB9-C1 represent the clinical backgrounds of patients with GCTB. Both cell lines were derived from patients in their thirties, aligning with the age commonly associated with this tumor [5]. The tumor origin sites, proximal and distal tibia, are epiphyses of the long bone and are frequent sites of GCTB occurrence [1]. Furthermore, the presence of the H3-3A G34W mutation in both cell lines was consistent with the established genomic features of GCTB. This mutation is crucial for an accurate understanding of GCTB pathogenesis because the H3-3A G34W mutation has been reported to block normal RNA processing and chromatin regulation and promote tumorigenic processes in GCTB [29].

The NCC-GCTB8-C1 and NCC-GCTB9-C1 cells exhibited reproducible properties suitable for in vitro functional studies. These cell lines exhibited consistent proliferative and invasive characteristics, which are critical for determining the efficacy of antitumor agents in preclinical settings. NCC-GCTB9-C1 cells showed a higher proliferative capacity. While the average doubling time for previously established GCTB strains in our laboratory was 26 h, NCC-GCTB9-C1 showed a much shorter doubling time of 12 h [6, 21–24]. NCC-GCTB9-C1 cells may be valuable as a representative cell line for GCTB cases with high local proliferative ability. Moreover, these cell lines demonstrated spheroid-formation capabilities. They have significant value because 3-dimensional cell cultures are being utilized as reliable models for drug discovery and development [30, 31].

In our comprehensive drug-screening assay involving NCC-GCTB8-C1 and NCC-GCTB9-C1 cells, along with other previously established GCTB cell lines, distinct drugs were identified as potential therapeutic candidates for GCTB, of which the HDAC inhibitor romidepsin and the topoisomerase inhibitor mitoxantrone, in particular, showed low IC₅₀ values. HDAC regulates gene expression via histone acetylation regulation and is associated with various cancer types and abnormalities, suggesting that it plays a pivotal role in tumor progression. Romidepsin has been used for cutaneous and peripheral T-cell lymphomas, although it has not been approved for clinical use in bone and soft tissue tumors, including GCTB, by the US Food and Drug Administration (FDA) [32]. However, the efficacy of HDAC inhibitors against GCTB, both in vitro and in vivo, has been previously reported in a preclinical setting, and our results are consistent with those studies [33, 34]. Mitoxantrone, a topoisomerase inhibitor, was designed to enhance the therapeutic properties of anthracyclines and is widely used in the treatment of breast and prostate cancers, lymphomas, and leukemias [35]. However, the efficacy of romidepsin and mitoxantrone in GCTB has not yet been established, indicating a need for further studies.

Our study has several limitations. Although this is the largest drug screening analysis conducted for GCTB, the number of cell lines and drugs used were insufficient compared with that used for common cancers. Additionally, the tumorigenic potential of these cell lines remains unclear. Thus, GCTB cell line behavior still has uncertainties in vivo, which need to be addressed for a more detailed understanding. Moreover, all GCTB cell lines established in this study were derived from primary lesions. Hence, they are not representative of recurrent or metastatic lesions, which are often clinically problematic. These challenges need to be addressed in future studies.

Conclusively, we established two novel GCTB cell lines, NCC-GCTB8-C1 and NCC-GCTB9-C1. These cell lines provide a robust basis for GCTB research and advance our understanding of the mechanisms underlying GCTB and potential therapeutic strategies. Our drug screening results highlight romidepsin and mitoxantrone as promising candidates for GCTB treatment. These findings encourage further investigation and validation in clinical settings to improve therapeutic outcomes in patients with GCTB.

Funding

This study was supported by the Japan Agency for Medical Research and Development (grant number: 20ck0106537h).

Conflicts of Interest

The authors have no relevant financial or non-financial interests to disclose.

Ethics approval

The use of clinical materials for this study was approved by the ethics committee of the National Cancer Center (approval number: 2004-050).

Informed consent

Written informed consent was provided by the patients.

Acknowledgments

We thank Drs. S. Fukushima, S. Osaki, S. Iwata (Department of Musculoskeletal Oncology and Rehabilitation), Y. Takahashi (Department of Diagnostic Pathology), and the National Cancer Center Hospital for sampling the tumor tissue specimens from surgically resected materials. We appreciate the technical assistance provided by Mr. Y. Ohno (Division of Rare Cancer Research, National Cancer Center Research Institute) and Mrs. Y. Shiotani (Central Animal Division, National Cancer Center Research Institute). We would like to thank Editage (www.editage.jp) for providing English language editing services and for their constructive comments on the manuscript. Technical assistance for this study was provided by the Fundamental Innovative Oncology Core of the National Cancer Center.

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Table 2 is available in the Supplementary Files section.

Table2IC50valuesGCTB89.xlsx
Table 2 Summary of half-maximal inhibitory concentration (IC₅₀) values in GCTB cell lines.
SupplementaryFigure1STR.tif
Supplementary Fig. 1 Short tandem repeat patterns of the cell lines and the original tumor tissue Short tandem repeat patterns of (A) original tumor tissue and (B) the cell line of NCC-GCTB8-C1. Short tandem repeat patterns of (C) the original tumor tissue and (D) the cell line of NCC-GCTB9-C1.
SupplementaryTable1Celllinelist.xlsx
Supplementary Table 1 List of existing GCTB cell lines registered in the Cellosaurus cell line database
SupplementaryTable2214drugslist.xlsx
Supplementary Table 2 List of the 214 anticancer drugs used in this study
SupplementaryTable3HTScellviabilityGCTB19.xlsx
Supplementary Table 3 Cell viability (%) after treatment with 214 anticancer drugs
SupplementaryTable4Drugtypeandclassification.xlsx
Supplementary Table 4 Drug type and classification
SupplementaryTable5IC50values.xlsx
Supplementary Table 5 IC₅₀ values

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published 11 Mar, 2024

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Editorial decision: Major Revisions Needed
01 Dec, 2023
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Establishment and characterization of two novel patient-derived cell lines from giant cell tumor of bone: NCC-GCTB8-C1 and NCC-GCTB9-C1

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Journal Publication

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Abstract

Figures

1. Introduction

2. Materials and methods

2.1 Patient data

2.2 Histological analysis

2.3 Establishment of cell lines

2.4 Authentication and quality control of the cell lines

2.5 Genetic analysis

2.6 Inspection of Mycoplasma contamination

2.7 Cell proliferation assay

2.8 Spheroid formation assay

2.9 Invasion assay

2.10 Drug screening assay

3. Results

3.1 Authentication of the established GCTB cell lines

3.2 Characterization of the established GCTB cell lines

3.3 Sensitivity of the antitumor agents to the established GCTB cell lines

4. Discussion

Declarations

References

Table 2

Supplementary Files

Status:

Journal Publication

Version 1