Murine hepatocyte cell lines promote expansion and differentiation of NK cells from stem cell precursors

Bordoni, Veronica; Alonzi, Tonino; Agrati, Chiara; Poccia, Fabrizio; Borsellino, Giovanna; Mancino, Giorgio; Fimia, Gian María; Piacentini, Mauro; Fantoni, A; Tripodi, Marco

doi:10.1002/hep.20234

Cited by 16 publications

(16 citation statements)

References 51 publications

(57 reference statements)

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“…Moreover, MMH cells express SCF, Flt3 ligand, IL-3, LIF, GM-CSF, G-CSF, TPO, IL-15, and IL-7. 24,25 The biological role of each of these factors remains uncharacterized; however, it is worth noting that MMH-CM induced morphological and functional differentiation toward endothelial cells only from CD34ϩ cells cultured as bulk populations and not as single cells. This result suggests that endothelial differentiation is also influenced by CD34ϩ autocrine/paracrine release of soluble factors.…”

Section: Discussionmentioning

confidence: 99%

“…MMH-conditioned media from embryonic E14 (MMH-CM) or from adult D6 (MMH-CMD6) MMH lines were obtained as previously described. 25 Briefly, semiconfluent (70%) cultures were washed with 1ϫ PBS, and the RPMI complete medium was replaced by IMDM, supplemented with 10% FBS and antibiotics without the addition of cytokines. After 48 hours, the medium was filtered through a 0.2-m filter and considered MMH-CM.…”

Section: Methodsmentioning

confidence: 99%

“…Flow cytometry was performed as described. 25 Capillary Tube Formation Assay. Capillary tube formation was assessed as described 31 using 5 ϫ 10 4 /well CD34-CM cells cultured for 21 days.…”

Section: Methodsmentioning

confidence: 99%

“…25 In this work we found that a MMH-conditioned medium (MMH-CM) maintains human umbilical cord blood (UCB)-derived CD34ϩ cells (CD34-CM) in culture for several weeks and directs their differentiation toward an endothelial lineage. Phenotypic and functional analyses showed that CD34ϩ cells gradually lose their hematopoietic potential when acquiring endothelial characteristics.…”

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confidence: 99%

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Hepatocyte-conditioned medium sustains endothelial differentiation of human hematopoietic-endothelial progenitors

et al. 2007

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Liver neo-angiogenesis plays a fundamental role in physiological and pathological processes such as regeneration, cirrhosis, autoimmune hepatitis, and alcoholic liver disease. How liver parenchymal cells influence angiogenesis is largely unknown. We studied the influence of soluble factors released by hepatocytes on hematopoietic and endothelial cell differentiation. Human CD34؉ cells cultured for several weeks in a hepatocyte-conditioned medium gradually decrease the expression of CD34 and CD133 markers (i.e. after 4 weeks from 85% and 69%, respectively, to 6% and 3%, respectively), whereas expression of CD144 and CD14 cell markers increased (from 2% and 8%, respectively, to 54% and 55%, respectively). The cells' capacity to form hematopoietic colonies in methylcellulose declined with time, whereas they acquired endothelial morphology, expressed endothelial markers, and incorporated into newly forming vascular structures both in vitro and in vivo. Cultured single CD34؉ cells formed colonies expressing both hematopoietic (CD45؉) and endothelial (CD144؉) markers, suggesting they constitute a bona fide hemangioblast population. Conclusion: This system allowed subsequent stages of differentiation of hematopoietic cells to endothelial cells to be defined, underlining the strict interrelationship between endothelial and hematopoietic cells in a hepatocyte environment.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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Hepatocyte-conditioned medium sustains endothelial differentiation of human hematopoietic-endothelial progenitors

et al. 2007

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“…In order to conduct an efficient in vitro analysis, hepatocyte isolates must be completely established. Although some methods for the isolation and identification of hepatocytes have been proposed to date (Crosby et al, 2001;Azuma et al, 2003;Bordoni et al, 2004;Tsuchiya et al, 2005), the purity, activity, and yield of these isolated hepatocytes are relatively low, and its mRNA integrity is much poorer (Nagaki et al, 2001;Morsiani et al, 2002). In order to further improve upon and solve these problems, in this study, the two-step collagenase perfusion technique was used to isolate hepatocytes on the basis of previous research.…”

Section: Introductionmentioning

confidence: 99%

Isolation and identification of bovine primary hepatocytes

Jiang

Li²,

Liu³

et al. 2013

Genet. Mol. Res.

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ABSTRACT. The liver is a unique organ that is endowed with a plethora of specialized functions. Most of its functional traits are controlled by hepatocytes. Primary hepatocytes have been used widely in in vitro models to understand the biological processes occurring in the liver. There are a number of methods used to separate hepatocytes, but the cell activity and purity are much lower in this condition. On the basis of previous research, in this study, the two-step collagenase perfusion technique was used for isolating hepatocytes. The key proteins of hepatocytes, cytokeratin-18 (CK-18) and albumin (ALB), were used to identify cells, and their contents were evaluated by immunohistochemistry and Western blotting. The results showed that the isolated hepatocytes comprised more than 96% of the corresponding protein volume stability. Therefore, this method was demonstrated to be reliable for identifying hepatocytes.

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