A reliable and economical method for gaining mouse embryonic fibroblasts capable of preparing feeder layers

Jiang, Guangming; Wan, Xin; Wang, Ming; Zhou, Jinlin; Pan, Jian; Wang, Baolong

doi:10.1007/s10616-014-9815-z

Cited by 5 publications

(5 citation statements)

References 30 publications

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“…Mouse embryonic fibroblasts (MEFs) from the E14.5 mouse embryos were isolated under sterile conditions as described previously (Jiang et al 2016). Briefly, mouse embryos were dissected, and then the head and internal organs were removed.…”

Section: Isolation and Treatment Of Mefsmentioning

confidence: 99%

Direct and indirect coculture of mouse hepatic progenitor cells with mouse embryonic fibroblasts for the generation of hepatocytes and cholangiocytes

et al. 2019

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The widespread use of hepatocytes and cholangiocytes for regenerative medicine and tissue engineering is restricted by the limited number of hepatocytes and cholangiocytes; a simple and effective method for the expansion and differentiation of the hepatic progenitor cells (HPCs) is required. Recent studies demonstrated that mouse embryonic fibroblasts (MEFs) play an important role in supporting the proliferation of the mouse hepatic progenitor cells (mHPCs). However, the effect of direct and indirect coculture of MEFs with mHPCs on the differentiation of mHPCs is poorly studied. Herein, we show that mHPCs rapidly proliferate and form colonies in direct or indirect contact coculture with MEFs in the serumfree medium. Importantly, after direct contact coculture of the mHPCs with MEFs for 6 days, mHPCs expressed the hepatic marker albumin (ALB) and did not express the cholangiocyte marker CK19, indicating their differentiation into hepatocytes. In contrast, after indirect contact coculture of the mHPCs with MEFs for 6 days, mHPCs expressed the cholangiocyte marker CK19 and did not express the hepatic marker ALB, indicating their differentiation into cholangiocytes. These results indicate that direct and indirect contact cocultures of the mHPCs with MEFs are useful for rapidly producing hepatocytes and cholangiocytes.

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Section: Isolation and Treatment Of Mefsmentioning

confidence: 99%

Direct and indirect coculture of mouse hepatic progenitor cells with mouse embryonic fibroblasts for the generation of hepatocytes and cholangiocytes

et al. 2019

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“…Mouse embryonic fibroblasts (MEFs) were isolated from E14.5 mouse embryos as described previously (Jiang et al, 2016). Briefly, mouse embryos were dissected and the head and internal organs were completely removed.…”

Section: Isolation and Treatment Of Mefsmentioning

confidence: 99%

TGFβ signaling controls intrahepatic bile duct development may through regulating the Jagged1‐Notch‐Sox9 signaling axis

Wang

Feng

Yasen

et al. 2018

Journal Cellular Physiology

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Due to the inherent limitations of the mouse models, the molecular mechanism of TGFβ signaling involved in the development of intrahepatic bile ducts (IHBDs) has been investigated little. Here, we investigated the role of TGFβ signaling and its regulatory mechanism in IHBDs development. We demonstrate that TGFβ signaling pathway activity is essential for IHBDs development. When blocking TGFβ signaling at E10.5, the number of bile ducts in hilum was reduced more than two fold and number of CK19 positive chlangiocytes in periphery was reduced more than 3.5-fold compared with controls. We also show that alpha-smooth muscle actin (α-SMA)-immunoreactive cells are located in the portal vein mesenchyme (PVM) adjacent to the bile ducts during IHBDs development and identify the α-SMA positive cells expressing the Notch ligand Jagged1 in the periportal area. Importantly, after blocking TGFβ signaling, the expression of Jagged1 was selectively decreased in the PVM but not in biliary epithelial cells (BECs), which is associated with the transformation of portal mesenchyme cells (PMCs) into portal myofibroblasts (PMFs). In addition, Sox9, which is downstream of Notch, is decreased after blocking the TGFβ signaling pathway in the liver. Our findings uncover a novel mechanism of TGFβ signaling in controlling the development of IHBDs may through regulating the Jagged1-Notch-Sox9 signaling axis.

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“…Mouse embryonic fibroblast (MEF) feeder cells were isolated and cultured as previously described. 30 Tissue samples were washed with sterile prewarmed phosphate-buffered saline solution (PBS, containing penicillin (100 U/mL) and streptomycin (0.1 mg/L). Fatty and connective tissues were then manually removed.…”

Section: Gastric Tissue Processing and Cell Culturementioning

confidence: 99%

A simple and cost‐efficient adherent culture platform for human gastric primary cells, as an in vitro model for Helicobacter pylori infection

et al. 2018

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A reliable and economical method for gaining mouse embryonic fibroblasts capable of preparing feeder layers

Cited by 5 publications

References 30 publications

Direct and indirect coculture of mouse hepatic progenitor cells with mouse embryonic fibroblasts for the generation of hepatocytes and cholangiocytes

Direct and indirect coculture of mouse hepatic progenitor cells with mouse embryonic fibroblasts for the generation of hepatocytes and cholangiocytes

TGFβ signaling controls intrahepatic bile duct development may through regulating the Jagged1‐Notch‐Sox9 signaling axis

A simple and cost‐efficient adherent culture platform for human gastric primary cells, as an in vitro model for Helicobacter pylori infection

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