1.Simple Maturation of Direct-Converted Hepatocytes Derived from Fibroblasts.
Young duck CHO ; Sangtae YOON ; Kyojin KANG ; Yohan KIM ; Seung Bum LEE ; Daekwan SEO ; Kiyoung RYU ; Jaemin JEONG ; Dongho CHOI
Tissue Engineering and Regenerative Medicine 2017;14(5):579-586
Target cells differentiation techniques from stem cells are developed rapidly. Recently, direct conversion techniques are introduced in various categories. Unlike pluripotent stem cells, this technique enables direct differentiation into the other cell types such as neurons, cardiomyocytes, insulin-producing cells, and hepatocytes without going through the pluripotent stage. However, the function of these converted cells reserve an immature phenotype. Therefore, we modified the culture conditions of mouse direct converted hepatocytes (miHeps) to mature fetal characteristics, such as higher AFP and lower albumin (ALB) expression than primary hepatocytes. First, we generate miHeps from mouse embryonic fibroblasts (MEFs) with two transcription factors HNF4α and Foxa3. These cells indicate typical epithelial morphology and express hepatic proteins. To mature hepatic function, DMSO is treated during culture time for more than 7 days. After maturation, miHeps showed features of maturation such as exhibiting typical hepatocyte-like morphology, increased up-regulated ALB and CYP enzyme gene expression, down-regulated AFP expressions, and acquired hepatic function over time. Thus, our data provides a simple method to mature direct converted hepatocytes functionally and these cells enable them to move closer to generating functional hepatocytes.
Animals
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Dimethyl Sulfoxide
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Fibroblasts*
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Gene Expression
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Hepatocytes*
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Methods
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Mice
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Myocytes, Cardiac
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Neurons
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Phenotype
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Pluripotent Stem Cells
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Stem Cells
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Transcription Factors
2.Stem cell analysis with intraductally treated mammary epithelial cells
Ah Young YOON ; Yun Young KIM ; Joon Hyop LEE ; Yoo Seung CHUNG ; Sangtae CHOI ; Jin Mo KANG ; Heung Kyu PARK ; Yong Soon CHUN
Korean Journal of Clinical Oncology 2019;15(1):27-33
PURPOSE: The activity of mammary stem cells (MaSCs) is essential to mammary growth, differentiation and regeneration in cycles of pregnancy, lactation, and involution. The capability to recruit the mammary gland through the cycles is attributed to stem cells. It was shown that the intraductal (i.duc) injection of pegylated liposomal doxorubicin (PLD) to multiparous FVB/N mice was associated with a significantly reduced outgrowth potential of mammary gland cells. We have explored i.duc PLD's effect on stem cell number and function in mouse mammary gland and aldehyde dehydrogenase (ALDH)'s availability as a mouse MaSC marker.METHODS: The total mammary epithelium was purified from 6 to 8-month-old FVB/N control and i.duc PLD-administered mice treated twice and analyzed by flow cytometry and limiting dilution cleared mammary fat pad transplants.RESULTS: There was no significant difference in the proportions of stem cell-enriched population (CD49(fhigh)CD24(med)) between control and i.duc PLD-treated groups. However, we found a significant reduction in the outgrowth potential of CD49(fhigh)CD24(med) and CD49(fhigh)CD24(med)ALDH(+) cells from i.duc PLD-treated mammary glands. We discovered that adding ALDH to CD49(fhigh)CD24(med) had the possibility of better marker selection for MaSC of mice.CONCLUSION: We present i.duc administration of PLD to reduce MaSC function, but not the number; and ALDH activity may add further selection of MaSCs to CD49f CD24 in mouse mammary glands. Screening of chemotherapeutic drugs or other natural products by this method of stem cell analysis may provide safe i.duc treatment in breast cancer.
Adipose Tissue
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Aldehyde Dehydrogenase
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Animals
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Biological Products
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Breast Neoplasms
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Doxorubicin
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Epithelial Cells
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Epithelium
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Female
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Flow Cytometry
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Humans
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Infant
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Lactation
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Mammary Glands, Human
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Mass Screening
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Methods
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Mice
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Pregnancy
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Regeneration
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Stem Cells