Related Experiment Video
Updated: Jul 4, 2025
Live-3D-Cell Immunocytochemistry Assays of Pediatric Diffuse Midline Glioma
Published on: November 11, 2021
[Is it possible to detect surface antigen CD133 on patient-derived glioblastoma continuous cell cultures using
V L Moiseenko1, O M Antipova1, S A Pavlova2
1Lomonosov Moscow State University, Moscow, Russia.
Abstract:
Theranostics combines diagnostics and therapeutic exposure. Regarding glioblastomas, theranostics solves the problem of detecting and destroying tumor stem cells resistant to irradiation and chemotherapy and causing tumor recurrence. Transmembrane surface antigen CD133 is considered as a potential marker of tumor stem cells.
Objective:
To detect CD133 in patient-derived glioblastoma continuous cell cultures using fluorescence microscopy and modified aptamers (molecular recognition elements) anti-CD133.
Material And Methods:
To detect CD133, we used mousey fluorescence monoclonal antibodies anti-CD133 MA1-219, FAM-modified DNA aptamers anti-CD133 AP-1-M and Cs5. Non-aptamer DNA oligonucleotide NADO was used as a negative control. Detection was performed for three samples of patient-derived glioblastoma continuous cell cultures coded as 1548, 1721 and 1793.
Results:
MA1-219 antibodies brightly stained cell culture 1548, to a lesser extent - 1721. There was diffuse staining of cell culture 1793. Cs5-FAM aptamer stained cells in a similar way, but much weaker. AP-1-M-FAM aptamer interacted with cells even weaker and diffusely stained only cell culture 1793. Non-aptamer NADO did not stain cell culture 1548 and very weakly diffusely stained cell culture 1793.
Conclusion:
For both molecular recognition elements (MA1-219 antibody and Cs5 aptamer), 3 cell culture samples can be arranged in the following order possibly reflecting CD133 status decrease: strong signal for cell culture 1548, much weaker for 1721, even weaker for 1793. Only cell culture 1548 can be considered CD133 positive with combination of Cs5+ and NADO signals. Cell culture 1793 is CD133 false positive with combination of Cs5+ and NADO+ signals.
Insights
This study aimed to detect CD133, a potential marker for glioblastoma stem cells, in cell cultures. Results showed varying CD133 expression levels, with only one culture definitively identified as CD133 positive.
Area of Science:
- Biomedical science
- Molecular biology
- Oncology
Context:
- Glioblastomas are aggressive brain tumors with recurrent disease often linked to therapy-resistant tumor stem cells.
- Theranostics offers a dual approach to diagnose and treat cancers by combining imaging and therapy.
- CD133 is a transmembrane protein investigated as a potential marker for glioblastoma stem cells.
Purpose:
- To detect the expression of CD133 in patient-derived glioblastoma cell cultures.
- To evaluate the efficacy of modified DNA aptamers and monoclonal antibodies as diagnostic tools for CD133 detection.
- To differentiate between true and false positive CD133 expression in glioblastoma cell lines.
Summary:
- Three patient-derived glioblastoma cell cultures (1548, 1721, 1793) were analyzed for CD133 expression using fluorescence microscopy.
- Detection employed anti-CD133 monoclonal antibody (MA1-219) and two anti-CD133 DNA aptamers (AP-1-M, Cs5), with a non-aptamer oligonucleotide (NADO) as a control.
- MA1-219 and Cs5 aptamer showed strong signals in culture 1548, weaker in 1721, and diffuse in 1793. Only culture 1548 was confirmed CD133 positive, while 1793 showed false positivity.
Impact:
- This research contributes to the development of targeted theranostic strategies for glioblastoma.
- Accurate detection of CD133 can aid in identifying and targeting glioblastoma stem cells, potentially improving treatment outcomes.
- The findings highlight the importance of careful validation of diagnostic markers to avoid false positives in cancer research.
More Related Videos
12:16Patient Derived Cell Culture and Isolation of CD133+ Putative Cancer Stem Cells from Melanoma
Published on: March 13, 2013
10:28Flow Cytometry-based Drug Screening System for the Identification of Small Molecules That Promote Cellular Differentiation of Glioblastoma Stem Cells
Published on: January 10, 2018