A rational approach for cancer stem-like cell isolation and characterization using CD44 and prominin-1(CD133) as selection markers
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Yi-Jen Lee1, Chang-Cheng Wu2, Jhy-Wei Li3,4, Chien-Chih Ou5, Shih-Chung Hsu6, Hsiu-Hsueh Tseng7, Ming-Ching Kao8,9, Jah-Yao Liu10,11
1Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan
2Chief of Obstetrics and Gynecology, Tri-Service General Hospital Penghu Branch, Penghu, Taiwan
3Chief of Pathology, Da-Chien General Hospital, Miaoli, Taiwan
4Department of Rehabilitation science, Jente Junior College of Medicine, Nursing and Management, Miaoli, Taiwan
5Obstetrics and Gynecology, Tri-Service General Hospital, Taipei, Taiwan
6Medical Care and Management, Kang-Ning Junior College, Taipei, Taiwan
7Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan
8Department of Biological Science and Technology, China Medical University, Taichung, Taiwan
9Department of Biochemistry, National Defense Medical Center, Taipei, Taiwan
10Department of Obstetrics and Gynecology, National Defense Medical Center, Taipei, Taiwan
11Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan
Jah-Yao Liu, email: email@example.com
Keywords: cancer stem-like cells (CSLC), intraperitoneal enrichment, CD44 and CD133, SKOV3.PX1_133+44+, OVCAR3.PX1_133+44+
Received: April 13, 2015 Accepted: September 12, 2016 Published: September 17, 2016
The availability of adequate cancer stem cells or cancer stem-like cell (CSC) is important in cancer study. From ovarian cancer cell lines, SKOV3 and OVCAR3, we induced peritoneal ascites tumors in immunodeficient mice. Among the cells (SKOV3.PX1 and OVCAR3.PX1) from those tumors, we sorted both CD44 and CD133 positive cells (SKOV3.PX1_133+44+, OVCAR3.PX1_133+44+), which manifest the characteristics of self-renewal, multi-lineage differentiation, chemoresistance and tumorigenicity, those of cancer stem-like cells (CSLC). Intraperitoneal transplantation of these CD44 and CD133 positive cells resulted in poorer survival in the engrafted animals. Clinically, increased CD133 expression was found in moderately and poorly differentiated (grade II and III) ovarian serous cystadenocarcinomas. The ascites tumor cells from human ovarian cancers demonstrated more CD133 and CD44 expressions than those from primary ovarian or metastatic tumors and confer tumorigenicity in immunodeficient mice. Compared to their parental cells, the SKOV3.PX1_133+44+ and OVCAR3.PX1_133+44+ cells uniquely expressed 5 CD markers (CD97, CD104, CD107a, CD121a, and CD125). Among these markers, CD97, CD104, CD107a, and CD121a are significantly more expressed in the CD133+ and CD44+ double positive cells of human ovarian ascites tumor cells (Ascites_133+44+) than those from primary ovarian or metastatic tumors. The cancer stem-like cells were enriched from 3% to more than 70% after this manipulation. This intraperitoneal enrichment of cancer stem-like cells, from ovarian cancer cell lines or primary ovarian tumor, potentially provides an adequate amount of ovarian cancer stem-like cells for the ovarian cancer study and possibly benefits cancer therapy.
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