Magnetic Cell Separation Kit for Human CD44v9+ Cancer Stem Cell

Cosmo Bio

Catalog No.: CSR-CSC-SEP1

For the Enrichment of Cancer Stem Cells

This kit includes magnetic particles conjugated with a monoclonal antibody that binds with high affinity to the membrane antigen CD44v8-10, which is frequently overexpressed in cancer stem cells (CSCs). By using Magnetic-activated Cell Sorting (MACS) with this kit, cancer stem cells can be enriched.

Background

In recent years, it has become clear that cancer stem cells are the source of carcinogenesis, such as the formation of cancerous tissue, its metastasis, and recurrence. Therefore, it is considered necessary to eliminate cancer stem cells in order to eradicate cancer, and cancer stem cells are drawing attention as the ultimate target for cancer treatment. However, cancer stem cells are present in tissues and cell lines in extremely small amounts (less than a few percent), so it has been difficult to develop therapeutic drugs targeting them and study their functional analysis.

This kit includes anti-CD44v9 antibody. This is a monoclonal antibody that binds with high affinity to the membrane antigen CD44v8-10, which is frequently overexpressed in cancer stem cells (CSCs). By using a cell sorter or similar equipment, this antibody makes it possible to easily achieve the enrichment of CSCs, which was previously difficult. The enrichment of CSCs allows for the establishment of evaluation systems for CSCs, such as the 'sphere culture system' used for in vitro evaluation and the 'mouse lung metastasis model' used for in vivo evaluation. This facilitates the elucidation of the mechanisms of cancer initiation and metastasis, as well as the screening of therapeutic drugs targeting CSCs.

Features

• Can easily enrich cancer stem cells with magnetic particles bound with cancer stem cell marker antibody
• Collecting cancer stem cells by a simple operation using a magnetic stand without using FACS
• Separated cells can be used for flow cytometric analysis

Kit Contents

• anti-CD44v9 magnetic beads (RV3 beads)
• Binding buffer
• Wash buffer

Workflow

Experimental Example

Isolate cells expressing CD44v9-GFP stably and analyze by flow cytometry

Figure 1. Ratio of CD44v9- expressing cells in RV3 bead-positive cells Stable expressing cells in which CD44v9-GFP was introduced into 293F cells and parent cell line 293F cells were mixed at any ratio (A, B, C). Was isolated using this product. CD44v9 expressing cells were concentrated from 4.97% (A), 15.3% (B) and 55.7% (C) before separation to 86.4% (A), 97.1% (B) and 99.0% (C) after separation.

Separation of cancer cells and analysis by flow cytometry

Figure 2. Ratio of CD44v9-expressing cells in RV3 bead-positive cells RV3 bead-positive cells were stained with PE-labeled anti-rat IgG antibody and analyzed by flow cytometry. Also in the human prostate cancer cell line PC-3 (A) and the human pancreatic adenocarcinoma cell line AsPC-1 (B), the ratio of CD44v9-expressing cells was increased compared to that before the isolation.

Citations for Magnetic Cell Separation Kit for Human CD44v9+ Cancer Stem Cell – 20 Found

Aguilar-Chaparro, Mario Alejandro; Rivera-Pineda, Sonia Andrea; Hernández-Galdámez, Hury Viridiana; Piña-Vázquez, Carolina; Villa-Treviño, Saúl. The CD44std and CD44v9 subpopulations in non-tumorigenic invasive SNU-423 cells present different features of cancer stem cells. Stem Cell Research. 2023;72( 37844417):103222. 

Mario Alejandro Aguilar-Chaparro 1, Sonia Andrea Rivera-Pineda 1, Hury Viridiana Hernández-Galdámez 1, Emmanuel Ríos-Castro 2, Olga Lilia Garibay-Cerdenares 3, Carolina Piña-Vázquez 1, Saúl Villa-Treviño 1. Transforming Growth Factor-β Modulates Cancer Stem Cell Traits on CD44 Subpopulations in Hepatocellular Carcinoma. J Cell Biochem . . 2025;126(2) 

Anna Kakehashi 1, Shugo Suzuki 1, Yusaku Nishidoi 1, Atsushi Hagihara 2, Hiroko Ikenaga 2, Masayuki Shiota 3, Guiyu Qiu 1, Ikue Noura 1, Yuko Kuwae 4, Arpamas Vachiraarunwong 1 5, Masaki Fujioka 1, Min Gi 1 5, Norifumi Kawada 2, Hideki Wanibuchi 1 5. The Impact of Peroxiredoxin 3 on Molecular Testing, Diagnosis, and Prognosis in Human Pancreatic Ductal Adenocarcinoma. Cancers (Basel).. 2025;17(13):2212. 

Tomona Yukimura 1, Tomohiro Seki 1, Toshinobu Seki 1. Preparation and Characterization of a Glutathione-Responsive Doxorubicin Prodrug Modified by 2-Nitrobenzenesulfonamide Group-Its Selective Cytotoxicity Toward Cells with Enhanced Glutathione Production. Int J Mol Sci. . 2025; 

Naoko Sasaki 1, Yoshiya Asano 2, Yukiko Sorayama 3, Chihiro Kamimura 3, Shiro Kitano 4, Shinji Irie 4, Ryohei Katayama 5, Hiroshi Shimoda 6, Michiya Matsusaki 7. Promoting biological similarity by collagen microfibers in 3D colorectal cancer-stromal tissue: Replicating mechanical properties and cancer stem cell markers. Acta Biomater. 2024;185:161-172. 

Zhibo Ma 1, Nikki K Lytle 1, Bob Chen 2, Nidhi Jyotsana 3, Sammy Weiser Novak 4, Charles J Cho 5, Leah Caplan 3, Olivia Ben-Levy 3, Abigail C Neininger 3, Dylan T Burnette 6, Vincent Q Trinh 7, Marcus C B Tan 8, Emilee A Patterson 9, Rafael Arrojo E Drigo 10, Rajshekhar R Giraddi 1, Cynthia Ramos 1, Anna L Means 11, Ichiro Matsumoto 12, Uri Manor 4, Jason C Mills 5, James R Goldenring 13, Ken S Lau 14, Geoffrey M Wahl 1, Kathleen E DelGiorno 15. Single-Cell Transcriptomics Reveals a Conserved Metaplasia Program in Pancreatic Injury. Gastroenterology. 2002;162(2) 

Jun Inoue 1, Masahiro Kishikawa 1 2, Hitoshi Tsuda 3, Yasuaki Nakajima 4, Takahiro Asakage 2, Johji Inazawa 1 5. Identification of PDHX as a metabolic target for esophageal squamous cell carcinoma. Cancer Sci. 2021;112(7):2792-2802. 

Yuka Okamoto 1, Takuya Saito 1, Yuri Tani 1, Tamami Toki 1, Akiko Hasebe 1, Masaru Koido 1, Akihiro Tomida 2. The kinase PERK represses translation of the G-protein-coupled receptor LGR5 and receptor tyrosine kinase ERBB3 during ER stress in cancer cells. J Biol Chem.. 2020;295(14)::4591-4603. 

Mariko Yasui 1 2, Akiko Kunita 1, Satoe Numakura 1 2, Hiroshi Uozaki 2, Tetsuo Ushiku 1, Masashi Fukayama 1 3. Cancer stem cells in Epstein-Barr virus-associated gastric carcinoma. Cancer Sci. 2020;111(7):2598-2607. 

Yamin Qian 1, Xin Wu 1, Yuhki Yokoyama 1, Daisuke Okuzaki 2, Mai Taguchi 1, Haruka Hirose 1, Jiaqi Wang 1, Tsuyoshi Hata 1 3, Akira Inoue 1 3, Masayuki Hiraki 1 3, Masahisa Ohtsuka 1 3, Hidekazu Takahashi 3, Naotsugu Haraguchi 3, Tsunekazu Mizushima 3, Shinji Tanaka 4, Masaki Mori 3 5, Hirofumi Yamamoto 1 3. E-cadherin-Fc chimera protein matrix enhances cancer stem-like properties and induces mesenchymal features in colon cancer cells. Cancer Sci. 2019;110(11):3520-3532. 

Tetsuo Mashima 1, Risa Iwasaki 2 3, Naomi Kawata 2 4, Ryuhei Kawakami 2 3, Koshi Kumagai 5, Toshiro Migita 2, Takeshi Sano 5, Kensei Yamaguchi 4, Hiroyuki Seimiya 6 7. In silico chemical screening identifies epidermal growth factor receptor as a therapeutic target of drug-tolerant CD44v9-positive gastric cancer cells. Br J Cancer.. 2019;121(10) 

Fumitaka Wada 1 2, Hironori Koga 1 2, Jun Akiba 3, Takashi Niizeki 1, Hideki Iwamoto 1 2, Yu Ikezono 1 2, Toru Nakamura 1 2, Mitsuhiko Abe 1 2, Atsutaka Masuda 1 2, Takahiko Sakaue 1 2, Toshimitsu Tanaka 1 2, Tatsuyuki Kakuma 4, Hirohisa Yano 3, Takuji Torimura 1 2. High expression of CD44v9 and xCT in chemoresistant hepatocellular carcinoma: Potential targets by sulfasalazine. Cancer Sci.. 2018;109(9):2801-2810. 

Masayuki Hagiwara 1, Eiji Kikuchi 2, Nobuyuki Tanaka 1, Takeo Kosaka 1, Shuji Mikami 3, Hideyuki Saya 4, Mototsugu Oya 1. Variant isoforms of CD44 involves acquisition of chemoresistance to cisplatin and has potential as a novel indicator for identifying a cisplatin-resistant population in urothelial cancer. BMC Cancer . 2018;18(1):113. 

Nattawan Suwannakul 1, Ning Ma 2, Raynoo Thanan 3 4, Somchai Pinlaor 4 5, Piti Ungarreevittaya 4 6, Kaoru Midorikawa 1, Yusuke Hiraku 1, Shinji Oikawa 1, Shosuke Kawanishi 7, Mariko Murata 1. Overexpression of CD44 Variant 9: A Novel Cancer Stem Cell Marker in Human Cholangiocarcinoma in Relation to Inflammation. Mediators Inflamm. 2018;2018(4867234) 

Megan D Radyk 1, Joseph Burclaff 1, Spencer G Willet 1, Jason C Mills 2. Metaplastic Cells in the Stomach Arise, Independently of Stem Cells, via Dedifferentiation or Transdifferentiation of Chief Cells. Gastroenterology. 2018;154(4):839-843. 

Chiara Di Stefano 1, Paola Grazioli 2, Rosaria Anna Fontanella 1, Paola De Cesaris 3, Antonella D'Amore 1, Michele Regno 1, Donatella Starace 1, Fabrizio Padula 1, Micol Elena Fiori 4, Rita Canipari 1, Antonella Stoppacciaro 5, Margherita Pesce 5, Antonio Filippini 1, Antonio Francesco Campese 6, Elio Ziparo # 1, Anna Riccioli # 1. Stem-like and highly invasive prostate cancer cells expressing CD44v8-10 marker originate from CD44-negative cells. Oncotarget.. 2018;9(56):30905-30918. 

Yushi Yamakawa 1, Masatoshi Kusuhara, Masanori Terashima, Yusuke Kinugasa, Takashi Sugino, Masato Abe, Toru Mochizuki, Keiichi Hatakeyama, Kenjiro Kami, Ken Yamaguchi. CD44 variant 9 expression as a predictor for gastric cancer recurrence: immunohistochemical and metabolomic analysis of surgically resected tissues. Biomed Res.. 2017;38(1):41-52. 

Hirokazu Kodama 1, Satoshi Murata 1, Mitsuaki Ishida 2 3, Hiroshi Yamamoto 1, Tsuyoshi Yamaguchi 1, Sachiko Kaida 1, Tohru Miyake 1, Katsushi Takebayashi 1, Ryoji Kushima 2, Masaji Tani 1. Prognostic impact of CD44-positive cancer stem-like cells at the invasive front of gastric cancer. Br J Cancer. . 116(2):186-194. 

Takeshi Miwa 1, Takuya Nagata 1, Hirofumi Kojima 1, Shinichi Sekine 1, Tomoyuki Okumura 1. Isoform switch of CD44 induces different chemotactic and tumorigenic ability in gallbladder cancer. Int J Oncol.. 2017;51(3):771-780. 

Shin Kasai 1, Yoshiyuki Furuichi 1, Norie Ando 1, Keiko Kagami 1, Masako Abe 1, Takaya Nakane 1, Kumiko Goi 1, Takeshi Inukai 1, Sei Saitoh 2, Shinichi Ohno 2, Shogo Okazaki 3, Osamu Nagano 3, Hideyuki Saya 3, Kanji Sugita 1. Inflammatory mediator ultra-low-molecular-weight hyaluronan triggers necrosis of B-precursor leukemia cells with high surface CD44 expression. Cell Death Dis.. 2017;8(6)