Generation of An Induced Pluripotent Stem Cell Line from Human Liver Fibroblasts from A Patient with Combined Hepatocellular-Cholangiocarcinoma
Objective: Combined hepatocellular-cholangiocarcinoma (cHCC-CC) is a rare type of primary liver cancer with characteristics of both hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC). The pathogenesis of cHCCCC is poorly understood due to a shortage of suitable in vitro models. Due to scar...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Royan Institute (ACECR), Tehran
2022-03-01
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| Series: | Cell Journal |
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| Online Access: | https://celljournal.org/journal/article/fulltext/generation-of-an-induced-pluripotent-stem-cell-line-from-human-liver-fibroblasts-of-a-patient-with-combined-hepatocellular-cholangiocarcinoma.pdf |
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| author | Hyo-Suk Ahn Jae-Sung Ryu Jaeseo Lee Seon Ju Mun Yeon-Hwa Hong Yongbo Shin Kyung-Sook Chung Myung Jin Son |
| author_facet | Hyo-Suk Ahn Jae-Sung Ryu Jaeseo Lee Seon Ju Mun Yeon-Hwa Hong Yongbo Shin Kyung-Sook Chung Myung Jin Son |
| author_sort | Hyo-Suk Ahn |
| collection | DOAJ |
| description | Objective: Combined hepatocellular-cholangiocarcinoma (cHCC-CC) is a rare type of primary liver cancer with
characteristics of both hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC). The pathogenesis of cHCCCC
is poorly understood due to a shortage of suitable in vitro models. Due to scarce availability of human liver tissue,
induced pluripotent stem cells (iPSCs) are a useful alternative source to produce renewable liver cells. For use in the
development of liver pathology models, here we successfully developed and evaluated iPSCs from liver fibroblasts of
a patient with cHCC-CC.
Materials and Methods: In this experimental study, human liver fibroblasts (HLFs) were obtained from the liver biopsy of
a 69-year-old male patient with cHCC-CC and transduced with a retroviral cocktail that included four factors - OCT4, SOX2,
KLF4, and c-MYC (OSKM). Pluripotency of the iPSCs was determined by alkaline phosphatase (AP) staining, quantitative
real-time polymerase chain reaction (PCR), and immunofluorescence. We induced in vitro embryoid body (EB) formation
and performed an in vivo teratoma assay to confirm their differentiation capacity into the three germ layers.
Results: HLF iPSCs derived from the cHCC-CC patient displayed typical iPSC-like morphology and pluripotency
marker expression. The proficiency of the iPSCs to differentiate into three germ layers was assessed both in vitro and
in vivo. Compared to normal control iPSCs, differentiated HLF iPSCs showed increased expressions of HCC markers
alpha-fetoprotein (AFP) and Dickkopf-1 (DKK1) and the CC marker cytokeratin 7 (CK7), and a decreased expression
of the CC tumour suppressor SRY-related HMG-box 17 (SOX17).
Conclusion: We established HLF iPSCs using liver fibroblasts from a patient with cHCC-CC for the first time. The HLF
iPSCs maintained marker expression in the patient when differentiated into EBs. Therefore, HLF iPSCs may be a sustainable
cell source for modelling cHCC-CC and beneficial for understanding liver cancer pathology and developing therapies for
cHCC-CC treatment. |
| first_indexed | 2024-04-12T22:23:32Z |
| format | Article |
| id | doaj.art-00d53389845544ad870ed67297f606df |
| institution | Directory Open Access Journal |
| issn | 2228-5806 2228-5814 |
| language | English |
| last_indexed | 2024-04-12T22:23:32Z |
| publishDate | 2022-03-01 |
| publisher | Royan Institute (ACECR), Tehran |
| record_format | Article |
| series | Cell Journal |
| spelling | doaj.art-00d53389845544ad870ed67297f606df2022-12-22T03:14:15ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142022-03-0124313313910.22074/cellj.2022.7765Generation of An Induced Pluripotent Stem Cell Line from Human Liver Fibroblasts from A Patient with Combined Hepatocellular-CholangiocarcinomaHyo-Suk Ahn0Jae-Sung Ryu1Jaeseo Lee2Seon Ju Mun3Yeon-Hwa Hong4Yongbo Shin5Kyung-Sook Chung6Myung Jin Son7Stem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, Republic of KoreaStem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, Republic of KoreaStem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, Republic of KoreaStem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, Republic of KoreaStem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, Republic of KoreaStem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, Republic of KoreaDepartment of Functional Genomics, Korea University of Science and Technology (UST), 217 Gajungro, Yuseong-gu, Daejeon, Republic of KoreaDepartment of Functional Genomics, Korea University of Science and Technology (UST), 217 Gajungro, Yuseong-gu, Daejeon, Republic of KoreaObjective: Combined hepatocellular-cholangiocarcinoma (cHCC-CC) is a rare type of primary liver cancer with characteristics of both hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC). The pathogenesis of cHCCCC is poorly understood due to a shortage of suitable in vitro models. Due to scarce availability of human liver tissue, induced pluripotent stem cells (iPSCs) are a useful alternative source to produce renewable liver cells. For use in the development of liver pathology models, here we successfully developed and evaluated iPSCs from liver fibroblasts of a patient with cHCC-CC. Materials and Methods: In this experimental study, human liver fibroblasts (HLFs) were obtained from the liver biopsy of a 69-year-old male patient with cHCC-CC and transduced with a retroviral cocktail that included four factors - OCT4, SOX2, KLF4, and c-MYC (OSKM). Pluripotency of the iPSCs was determined by alkaline phosphatase (AP) staining, quantitative real-time polymerase chain reaction (PCR), and immunofluorescence. We induced in vitro embryoid body (EB) formation and performed an in vivo teratoma assay to confirm their differentiation capacity into the three germ layers. Results: HLF iPSCs derived from the cHCC-CC patient displayed typical iPSC-like morphology and pluripotency marker expression. The proficiency of the iPSCs to differentiate into three germ layers was assessed both in vitro and in vivo. Compared to normal control iPSCs, differentiated HLF iPSCs showed increased expressions of HCC markers alpha-fetoprotein (AFP) and Dickkopf-1 (DKK1) and the CC marker cytokeratin 7 (CK7), and a decreased expression of the CC tumour suppressor SRY-related HMG-box 17 (SOX17). Conclusion: We established HLF iPSCs using liver fibroblasts from a patient with cHCC-CC for the first time. The HLF iPSCs maintained marker expression in the patient when differentiated into EBs. Therefore, HLF iPSCs may be a sustainable cell source for modelling cHCC-CC and beneficial for understanding liver cancer pathology and developing therapies for cHCC-CC treatment.https://celljournal.org/journal/article/fulltext/generation-of-an-induced-pluripotent-stem-cell-line-from-human-liver-fibroblasts-of-a-patient-with-combined-hepatocellular-cholangiocarcinoma.pdfcholangiocarcinomahepatocellular carcinomainduced pluripotent stem cells |
| spellingShingle | Hyo-Suk Ahn Jae-Sung Ryu Jaeseo Lee Seon Ju Mun Yeon-Hwa Hong Yongbo Shin Kyung-Sook Chung Myung Jin Son Generation of An Induced Pluripotent Stem Cell Line from Human Liver Fibroblasts from A Patient with Combined Hepatocellular-Cholangiocarcinoma Cell Journal cholangiocarcinoma hepatocellular carcinoma induced pluripotent stem cells |
| title | Generation of An Induced Pluripotent Stem Cell Line from Human Liver Fibroblasts from A Patient with Combined Hepatocellular-Cholangiocarcinoma |
| title_full | Generation of An Induced Pluripotent Stem Cell Line from Human Liver Fibroblasts from A Patient with Combined Hepatocellular-Cholangiocarcinoma |
| title_fullStr | Generation of An Induced Pluripotent Stem Cell Line from Human Liver Fibroblasts from A Patient with Combined Hepatocellular-Cholangiocarcinoma |
| title_full_unstemmed | Generation of An Induced Pluripotent Stem Cell Line from Human Liver Fibroblasts from A Patient with Combined Hepatocellular-Cholangiocarcinoma |
| title_short | Generation of An Induced Pluripotent Stem Cell Line from Human Liver Fibroblasts from A Patient with Combined Hepatocellular-Cholangiocarcinoma |
| title_sort | generation of an induced pluripotent stem cell line from human liver fibroblasts from a patient with combined hepatocellular cholangiocarcinoma |
| topic | cholangiocarcinoma hepatocellular carcinoma induced pluripotent stem cells |
| url | https://celljournal.org/journal/article/fulltext/generation-of-an-induced-pluripotent-stem-cell-line-from-human-liver-fibroblasts-of-a-patient-with-combined-hepatocellular-cholangiocarcinoma.pdf |
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