Hematopoietic Cells Derived from Cancer Stem Cells Generated from Mouse Induced Pluripotent Stem Cells.

Ghmkin Hassan,Hager Mansour,Kazuki Kumon,Hagar A Abu Quora,Ayano Satoh,Masaharu Seno,Amira Osman,Neha Nair,Hend M Nawara,Akimasa Seno,Juan Du,Nobuhiro Okada,Said M Afify,Maram H Zahra

doi:10.3390/cancers12010082

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

Introduction

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.

Methods

Results

Discussion

Conclusion