The Construction and Identification of Induced Pluripotent Stem Cells Derived from Acute Myelogenous Leukemia Cells

Abstract

Objective: The present study aimed to establish an induced pluripotent stem cell (iPSC) line from acute myelogenous leukemia (AML) cells in vitro and identify their biological characteristics. Methods: Cells from the AML-infiltrated skin from an M6 patient were infected with a lentivirus carrying OCT4, SOX2, KLF4 and C-MYC to induce iPSCs. The characteristics of the iPSCs were confirmed by alkaline phosphatase (ALP) staining. The proliferation ability of iPSCs was detected with a CCK-8 assay. The expression of pluripotency markers was measured by immunostaining, and the expression of stem cell-related genes was detected by qRT-PCR; distortion during the induction process was detected by karyotype analysis; the differentiation potential of iPSCs was determined by embryoid body-formation and teratoma-formation assays. ALP staining confirmed that these cells exhibited positive staining and had the characteristics of iPSCs. Results: The CCK-8 assay showed that the iPSCs had the ability to proliferate. Immunostaining demonstrated that iPSC clones showed positive expression of NANOG, SSEA-3, SSEA-4, TRA-1-60 and TRA-1-81. qRT-PCR results revealed that the mRNA expression of Nanog, Lin28, Cripto, FOX3, DNMT3b, DPPA2, and DPPA4 significantly increased in iPSCs. Karyotype analysis found no chromosome aberration in the iPSCs. The results of the embryoid body-formation and teratoma-formation assays indicated that the iPSCs had the potential to differentiate into all three germ layers. Conclusion: Our study provided evidence that an iPSC line derived from AML cells was successfully established.

Cancer cells as a new source of induced pluripotent stem cells

“…Melanocyte-iPSC lines showed pluripotency markers and the ability to differentiate into embryoid bodies in vitro and teratoma formation after injection into the dorsal flank of mice in vivo [ 12 ]. There are some reports of cancer-iPSCs teratoma formation [ 13 – 15 , 25 ], pluripotency character [ 9 , 14 , 26 , 27 ], and differentiation capacity [ 26 ] (Fig. 1 ).…”

“…Malignant tumors are cancerous [ 40 ]. There are some reports of teratoma formation for cancer-iPSCs derived from Acute Myelogenous Leukemia Cells [ 13 ], breast cancer cells [ 14 ], prostate cancer cells [ 15 ], and osteosarcoma cells [ 25 ]. iPSCs can make teratomas quicker and more efficiently than ESCs in vivo, indicating their higher potency in making tumors [ 41 ].…”

Analysis of lncRNA-Associated ceRNA Network Reveals Potential lncRNA Biomarkers in Human Colon Adenocarcinoma

“…Finally, 42 mRNAs were found to interact with 18 miRNAs (Table 3). Among these mRNAs, some were validated to be cancer-related genes, such as TPM2, KLF4, and EPHA7 [10][11][12].…”

DPPA2, DPPA4, and other DPPA factor epigenomic functions in cell fate and cancer

“…DPPA2 and DPPA4 were tested for their ability to reprogram somatic cells to induced pluripotent stem cells (iPSCs) in the Yamanaka laboratory's screens due to their high and selective expression in ESCs ( Takahashi and Yamanaka, 2006 ). Neither DPPA2 nor DPPA4 made the final list of factors minimally required for reprogramming of somatic cells to iPSC; however, many studies continued to use them as markers of both pluripotency in general and successful reprogramming due to their strikingly selective expression pattern in pluripotent stem cells ( Hu et al., 2010 ; Zhu et al., 2017 ).…”