Abstract
Cancer stem cells (CSCs) represent the subpopulation of cancer cells with the ability to differentiate into other cell phenotypes and initiated tumorigenesis. Previously, we reported generating CSCs from mouse induced pluripotent stem cells (miPSCs). Here, we investigated the ability of the CSCs to differentiate into hematopoietic cells. First, the primary cells were isolated from malignant tumors that were formed by the CSCs. Non-adherent cells (NACs) that arose from adherent cells were collected and their viability, as well as the morphology and expression of hematopoietic cell markers, were analyzed. Moreover, NACs were injected into the tail vein of busulfan conditioned Balb/c nude mice. Finally, CSCs were induced to differentiate to macrophages while using IL3 and SCF. The round nucleated NACs were found to be viable, positive for hematopoietic lineage markers and CD34, and expressed hematopoietic markers, just like homing to the bone marrow. When NACs were injected into mice, Wright–Giemsa staining showed that the number of white blood cells got higher than those in the control mice after four weeks. CSCs also showed the ability to differentiate toward macrophages. CSCs were demonstrated to have the potential to provide progenies with hematopoietic markers, morphology, and homing ability to the bone marrow, which could give new insight into the tumor microenvironment according to the plasticity of CSCs.
Highlights
Cancers have complex architecture, in which the malignant cells interact with non-transformed cells, forming the tumor microenvironment (TME) [1]
Our lab has previously established a protocol for generating Cancer stem cells (CSCs) from mouse induced pluripotent stem cells (miPSCs) while using
BT549 cells was used to convert into CSCs, CSCcmBT549 cells [12]
Summary
In which the malignant cells interact with non-transformed cells, forming the tumor microenvironment (TME) [1]. The identification of different components of TME, to which the tumor cells respond, could help to understand the chemoresistance in more detail and contribute to the development of more effective therapy [2,3]. Blood vessels and immune cells are considered as the main components of the TME influencing the progression and growth of tumors. Two μg of pAcmCherry-C2 DNA was transfected to 2 × 106 CSCcmBT549 cells that were suspended in 600 μL of Gene Pulser electroporation buffer (Bio-Rad Laboratories, Hercules, CA, USA). The cell suspension was electroporated in 0.4-cm gap-cuvette by Gene Pulser II (Bio-Rad Laboratories, Hercules, CA, USA) and the cells were seeded in gelatin-coated dishes. MCherry- and GFP-positive undifferentiated cells were both further selected in the presence of 1 ug/mL puromycin (Sigma-Aldrich, St. Louis, MO, USA) for seven days.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
