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Liquid Biopsy

 

  • Real-time information on tumor progression
  • Cost-effective way to tailor the therapy
  • Increase of therapy efficiency
  • Reduction of drug side-effects

 

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Personalized cancer therapy using circulating tumor cells (CTCs)

  • Easy, fast and  inexpensive
  • No extra costs
  • All types of solid tumors – cell surface
  • Antigen-independent
  • Human and animal samples
  • Gentle to isolated cells – keeping CTCs and DTCs
  • Viable for cultivation and downstream analysis
Liquid Biopsy
Product Size Price
New MetaCell
EMC001
10 pcs/box
 
Summary

Summary

PRINCIPLE OF THE METHOD

The MetaCell® kit is intended for a simple separation of circulating rare cells from peripheral blood (or other body fluids). MetaCell® technology uses a size-based separation principle. The following cell types may be enriched: circulating tumor cells (CTCs), disseminated tumor cells (DTCs), circulating tumor microemboli (CTM) and circulating endometrial cells (CECs).

 

As a result of the gentle flow of blood during the separation process, the enriched CTCs/ DTCs/ CTM/ CECs are in a very good viable condition. It is therefore possible to prepare a short-term (up to 5-7 days) in vitro culture of the enriched cells on a standard separation polycarbonate membrane.

The separation procedures are designed to enhance prognosis. A patient’s response to treatment can be assessed by confirming the presence of circulating rare cells.  

Note: Before use, all components of the MetaCell separation kit should be incubated under the UV – light for a minimum of 15 minutes to prevent external contamination. Special caution is required if long-term in vitro culture is planned and a prolonged time of incubation under UV is recommended.

THE METACELL® TECHNOLOGY

SIMPLE:

MetaCell® uses a capillary–action driven size-based separation of rare blood cells from the unclotted peripheral blood. The MetaCell® enables separation and detection of rare cells within a single step. Approximately 8-10 ml of blood is passed through a MetaCell® tube; the collection of the CTCs/DTCs on the separating membrane takes 2 minutes. MetaCell® technology does not require any expensive equipment or additional investment. One test = one tube. Please add remark on microscope.

SENSITIVE:
MetaCell® is size-based specific to rare cells in peripheral blood without dependency on the tumor cell membrane antigens (EpCAM, HER2, MUC1, EGFR, etc.). MetaCell® technology is suitable for enrichment of rare cells spread from epithelial as well as mesenchymal solid tumors. Separation of circulating endometrial cells from blood of endometriosis patients is also effective.

SOFT:
MetaCell® is soft to the separated cells, keeping them viable. After the separation, the viable intact cells are suitable for subsequent characterization and/or in vitro cultivation. The cell separation and cultivation platform combines sophisticated size-based separation with cultivation on the membrane principle.

KIT COMPOSITION

MetaCell® separation kit consists of a filling funnel, a plastic ring holding a separation membrane and a blood reservoir containing blood absorbents.

 

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MetaCell® separation tube consists of filling funnel (1), which is accompanied by a plastic ring (2) holding a separation membrane (3), the tube and ring complex are fixed in a flown blood reservoir (4).

FEATURES

  • Enrichment of intact of CTCs/DTCs/circulating endometrial cells (CECs)/circulating tumor microemboli (CTM) in very short time (2-3 min)
  • No problems with flow-based blood coagulation
  • Isolation of both epithelial and mesenchymal tumor cells without use of antibodies against cell membrane antigens
  • High detection capability – 1 tumor cell in 8-10 ml of blood
  • Separation process does not require any use of lysis reagents: (a simple gentle filtration without affecting the cell character)
  • Preservation of the morphological attributes of isolated cells
  • Possibility to count isolated cells immediately after separation process
  • Possibility to culture enriched cells in vitro for subsequent use/analysis/characterization
  • No need for accessory equipment/ machine for cell separation, no space requirements


CTC Identification Process

CTC Identification Process

Cytomorphology + Molecular Diagnostics = CTC

Protocol – 30 min

Blood collection:
Approximately 8-10 ml of venous blood is drawn from the antecubital veins and placed into an EDTA-containing tube.

Size-based cell separation:
8 ml of peripheral blood sample (can be extended up to 50 ml) is transferred into the tube and filtered through an 8 μm-pore polycarbonate membrane.

In vitro culture set up and cytomorphological analysis:
The membrane is transferred into a 6-well cultivation plate and observed by inverted microscope.

 

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CTCs and DTCs cultivation and down stream analysis

Separated cells can be cultured and the grown cells can be analyzed by
  • histochemistry (e.g. MGG staining)
  • immunohistochemistry using specific antibodies to determine the cell origin (membrane is compatible with staining machines e.g. Ventana)
  • vital fluorescence staining, which enables to use stained cells for further downstream analysis can be used
  • gene expression profiling (mutational analysis, methylation analysis, tumor associated markers, therapy-related markers).

 

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CLINICAL INDICATIONS FOR CTC EXAMINATION / BREAST CANCER STUDY

Circulating tumor cell (CTC) examination was indicated as a complementary test during neoadjuvant therapy in 20 patients with breast cancer. The presented findings support the use of liquid biopsy to enhance monitoring of malignant disease and prediction of treatment efficacy. During the CTC-examination CTC presence was monitored. In parallel, gene expression analysis was performed on isolated enriched CTCs. Tumor associated genes and genes associated with chemoresistance were evaluated in comparison to the administered therapy (anthracyclines/taxanes/ anti-Her2therapy).
In general, elevated CTC-count, indicates chemoresistance, usually. However, timely retuning of therapy based on CTC-chemoresistance data obtained with the liquid biopsy resulted in reasonable decreases in tumor progression expressed as the CTC-count.

 

SITUATION 1: The standard neoadjuvant with anthracyclines (AC). Regardless of whether the tumor is reduced, the number of AC resistant CTCs increases (increased expression of MRP1); the treatment continues. The patient is indicated for surgical removal of the tumor. After tumor removal, the number of CTCs decreases for a while but then again rises with the formation of metastatic lesions.

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SITUATION 2: During the AC treatment the therapy is changed (based on CTC resistance) to taxanes. The tumor is reduced and the number of CTCs decreases. After the surgical removal, the tumor is reduced and CTCs decreases.

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What if the chemosensitivity test is performed before chemotherapy start?

SITUATION 3: At the very beginning of the treatment, presence of AC resistance on CTCs is determined, therefore the taxanes are indicated immediately. After the therapy start, tumor volume is reduced and the number of CTC decreases.

 

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Applications

Applications

The MetaCell® technology enables

  • CTC capture
  • CTC culture
  • CTC characterization

 

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The MetaCell® technology can be used in

Science: Principles of cancer dissemination may be observed in parallel to the primary tumor tissue and metastasis.

 

Health-care providers
Personalized therapy within precision medicine could save patients from side effects and enormous treatment costs. Detailed information on the characteristics cells enriched from circulation or body fluids enables the clinicians to select an appropriate treatment for their patients.

Pharma and biotech industry
Co-development of drug and companion diagnostics, discovery of new druggable markers (CTCs offer a new possible target for oncotherapy).

The use of the MetaCell® tests has been proved in

Oncology
Breast cancer, gastric cancer, lung cancer, esophageal cancer, pancreatic cancer, prostate cancer, urothelial tumors…


Gynecology
Circulating tumor cells (CTCs), circulating endometrial – like cells (CECs), circulating trophoblasts (CTLC), breast cancer, ovarian cancer, cervical cancer, endometrial cancer, endometriosis, pregnancy…

Animal studies
Orthotopic metastatic nude mice models (Human cancer lines are implanted in nude mice. Please see the references list.).

References to Product

References to Product

  • Kolostova, Katarina, et al. The added value of circulating tumor cells examination in ovarian cancer staging. American journal of cancer research 5.11 (2015): 3363.
  • Kolostova, Katarina, et al. Molecular characterization of circulating tumor cells in ovarian cancer. American journal of cancer research 6.5 (2016): 973.
  • Kolostova, Katarina, et al. Isolation, primary culture, morphological and molecular characterization of circulating tumor cells in gynecological cancers. American journal of translational research 7.7 (2015): 1203.
  • Kolostova, Katarina, et al. Morphological and gene-expression characterization of viable heterogeneous circulating tumor cells size-captured and cultured from triple-negative breast cancer mouse models. Int J Clin Exp Med 9.5 (2016): 7772-7779.
  • Kolostova, Katarina, et al. In vitro culture and characterization of human lung cancer circulating tumor cells isolated by size exclusion from an orthotopic nude-mouse model expressing fluorescent protein. Journal of fluorescence 24.5 (2014): 1531-1536.
  • Bobek, Vladimir, Katarina Kolostova, and Eduard Kucera. Circulating endometrial cells in peripheral blood. European Journal of Obstetrics & Gynecology and Reproductive Biology 181 (2014): 267-274.
  • Bobek, Vladimir, et al. Cultivation of circulating tumor cells in esophageal cancer. Folia Histochem Cytobiol 52.3 (2014): 171-177.
  • Kolostova, Katarina, et al. Detection and cultivation of circulating tumor cells in gastric cancer. Cytotechnology 68.4 (2016): 1095-1102.
  • Bobek, Vladimir, et al. Circulating tumor cells in pancreatic cancer patients: enrichment and cultivation. World Journal of Gastroenterology: WJG 20.45 (2014): 17163.
  • Bobek, Vladimir, Martin Cegan, and Katarina Kolostova. Circulating tumour cells in patients with urothelial tumours: Enrichment and in vitro culture. Canadian Urological Association Journal 8.9-10 (2014): 715-20.

 


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