Development of extracellular matrix supported 3D culture of renal cancer cells and renal cancer stem cells

Maliszewska‐Olejniczak, Kamila; Brodaczewska, Klaudia; Bielecka, Zofia F.; Solarek, Wojciech; Kornakiewicz, Anna; Szczylik, Cezary; Porta, Camillo; Czarnecka, Anna M.

doi:10.1007/s10616-018-0273-x

Cited by 18 publications

(13 citation statements)

References 34 publications

(55 reference statements)

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“…Since that time, a revolution in the methods and approach to cell culture has begun, and scientists learned how to isolate and culture cells from different tissues [ 15 ]. However, researchers observed that in some cases, the two-dimensional (2D) cell culture does not answer the questions about the structure of the tissue, and cell to cell interactions, and therefore the effect of the agents tested can’t be properly analyzed [ 16 ]. The better solution would be the preparation of the 3D constructs but this is difficult.…”

Section: Discussionmentioning

confidence: 99%

A Simple Method for the Production of Human Skin Equivalent in 3D, Multi-Cell Culture

Szymański

Jęderka

Cios

et al. 2020

IJMS

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An important problem for researchers working in the field of dermatology is the preparation of the human skin equivalent (HSE). Here, we describe a simple and reliable protocol for preparing a skin model from the commercially available cell lines: keratinocytes, fibroblasts, and melanocytes. Importantly, in our 3D model, the keratinocytes are diverse that brings this model closer to the natural skin. For the production of HSE, we used available primary PCS-200-010, PCS-201-010, PCS-200-013, and immortalized CRL-4048 and CRL-4001 cell lines. We used genipin, which is necessary for collagen cross-linking and studied its cytotoxicity for keratinocytes and fibroblasts. The addition of 20 μM genipin reduced the shrinkage of the collagen in the constructs from 59% to 24% on day 12 of the culture of the construct. A higher concentration (80–200 µM) of genipin reduced shrinkage by 14% on average. Genipin in concentration 10 μM and below was not cytotoxic to the keratinocytes, and 150 μM and below to the fibroblasts. Hematoxylin and eosin staining showed that the morphology of HSEs was identical to that of native human skin. The immunohistochemical staining of the constructs showed the presence of vimentin-positive fibroblasts in the skin layer, while the melanocytes were in the epidermis and in the basal layer. We observed that the longer differentiation of constructs led to the higher secretion of GM-CSF, IL-10, IL-15, IL-1α, IL-6, IL-7, IL-8, and MCP-1. We also observed that the longer time of differentiation led to a more stable secretion of all analytes, which was reflected in the coefficient of variation. We described here a simple, reliable, and cost-effective production of the full-thickness human skin equivalents that can be used in the research and industry. With the global trend to decrease animal use for the research and testing, our HSE could be a useful testing tool and an alternative research model.

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Section: Discussionmentioning

confidence: 99%

A Simple Method for the Production of Human Skin Equivalent in 3D, Multi-Cell Culture

Szymański

Jęderka

Cios

et al. 2020

IJMS

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“…The terms “sphere”, “tumor sphere” [ 25 ] and “oncosphere” [ 26 ] are used to describe the spheres obtained from CSCs. Tumor spheres are a mixture of CSCs and tumor cells, since CSCs are known for their ability to proliferate and differentiate [ 27 ]. This model is commonly used to evaluate and characterize CSCs in vitro [ 28 ].…”

Section: Tumor Spheroids and Organoidsmentioning

confidence: 99%

Promising Applications of Tumor Spheroids and Organoids for Personalized Medicine

Gilazieva

Ponomarev

Rutland

et al. 2020

Cancers

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One of the promising directions in personalized medicine is the use of three-dimensional (3D) tumor models such as spheroids and organoids. Spheroids and organoids are three-dimensional cultures of tumor cells that can be obtained from patient tissue and, using high-throughput personalized medicine methods, provide a suitable therapy for that patient. These 3D models can be obtained from most types of tumors, which provides opportunities for the creation of biobanks with appropriate patient materials that can be used to screen drugs and facilitate the development of therapeutic agents. It should be noted that the use of spheroids and organoids would expand the understanding of tumor biology and its microenvironment, help develop new in vitro platforms for drug testing and create new therapeutic strategies. In this review, we discuss 3D tumor spheroid and organoid models, their advantages and disadvantages, and evaluate their promising use in personalized medicine.

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“…As CD105−expressing subpopulation in human RCC xenografts from patient (nephrectomy) samples was shown to possess high capability of self-renewal and ability to form spheres in cell culture in vitro in complement to tumors in mice 28 it remains interesting that CD105− cells also give rise to tumors in our study. It is known that TGFβ pathway activation promote RCC 3D sphere formation and CD105 expression is up-regulated in stem-promoting conditions 33 . RCC cells derived spheres have actually been shown to exhibit cancer stem cell (tumor initiating cell) properties: self-renewal, tumorigenicity and the ability to differentiate into other cell types of original tumor 34 .…”

Section: Discussionmentioning

confidence: 99%

Renal carcinoma CD105−/CD44− cells display stem-like properties in vitro and form aggressive tumors in vivo

Fiedorowicz

Khan

Strzemecki

et al. 2020

Sci Rep

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clear cell renal cell carcinoma (ccRcc) is the most common kidney cancer. prognosis for ccRcc is generally poor since it is largely resistant to chemo-and radiotherapy. Many studies suggested that cancer stem cells/tumor initiating cells (CSCs/TICs) are responsible for development of tumor, disease progression, aggressiveness, metastasis and drug resistance. However, tumorigenic potential of CSCs/ tics isolated from established Rcc cell lines-basic ccRcc research model-has never been investigated in vivo. CD105+, CD105−, CD44+ and CD44− as well as CD44−/CD105− CD44+/CD105+ and CD44−/ CD105+ cells were isolated from Caki-1 RCC cell line, confirming coexistence of multiple subpopulations of stem-related phenotype in stable cell line. Sorted cells were injected subcutaneously into noD SciD mice and tumor growth was monitored with MRi and pet/ct. tumor growth was observed after implantation of CD105+, CD44+, CD44−, CD44−/CD105+ and CD44−/CD105− but not CD105− or CD44+/CD105+. Implantation of CD44−/CD105− cells induced tumors that were characterized by longer T1 and distinct metabolic pattern than other tumors. All the tumors were characterized by low uptake of [18F]FDG. CD105+ and CD44− tumors expresses Nanog and Oct-4, while CD44− tumors additionally expressed endothelial cell marker-CD31. Renal cell carcinoma (RCC), is the 10th malignancy worldwide and the most frequent type of kidney cancer in adults. Each year in Europe approximately 88 400 patients are diagnosed with RCC; the incidence and mortality of RCC are rising at a rate of 2-3% per decade, therefore novel therapies directed against RCC are needed. At the same time despite advancements in diagnostic techniques, up to 30% of newly diagnosed patients already present with metastases, and a large portion of patients that undergo surgical treatment experience the RCC recurrence, therefore drugs targeted against metastasis initiating cells would be of great interest in the future 1,2. Cancer stem cells (CSCs) are characterized by the potential to self-renew, high tumorigenicity in nude mice and the ability to efficiently reconstitute all tumor subpopulations and primary tumor phenotype 3-5. CSCs are also responsible not only for cancer development, but also for disease recurrence, progression and metastatic spread, together with cancer aggressiveness, including treatment resistance such as chemo/radiotherapy, and targeted treatment 6,7 , therefore basic research with careful model selection to understand their biology is mandatory to define novel potential therapeutic targets for all RCC subtypes 8,9 .

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Development of extracellular matrix supported 3D culture of renal cancer cells and renal cancer stem cells

Cited by 18 publications

References 34 publications

A Simple Method for the Production of Human Skin Equivalent in 3D, Multi-Cell Culture

A Simple Method for the Production of Human Skin Equivalent in 3D, Multi-Cell Culture

Promising Applications of Tumor Spheroids and Organoids for Personalized Medicine

Renal carcinoma CD105−/CD44− cells display stem-like properties in vitro and form aggressive tumors in vivo

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