Rat and mouse epiblasts differ in their capacity to generate extraembryonic endoderm

1998 ◽  
Vol 10 (8) ◽  
pp. 517 ◽  
Author(s):  
Jennifer Nichols ◽  
Austin Smith ◽  
Mia Buehr
Keyword(s):  
In Vitro Differentiation ◽  
Differentiation Potential ◽  
Rat Embryos ◽  
Extraembryonic Endoderm ◽  
Parietal Endoderm

In this study we have compared the in vitro differentiation potential of epiblast tissue from mouse and rat embryos. Epiblasts were isolated from egg cylinder stage embryos by microdissection and placed in culture. Rat cultures were distinguished by the copious production of parietal endoderm cells. Mouse epiblasts, in contrast, did not produce parietal endoderm. This difference in capacity to regenerate extraembryonic endoderm marks a surprising distinction in development of the pluripotential lineage between these two closely related rodents.

296 IN VITRO DIFFERENTIATION OF PORCINE BONE MARROW-DERIVED MESENCHYMAL STEM CELLS INTO HEPATOCYTE-LIKE CELLS

2013 ◽  
Vol 25 (1) ◽  
pp. 295
Author(s):  
B. Mohana Kumar ◽  
W. J. Lee ◽  
Y. M. Lee ◽  
R. Patil ◽  
S. L. Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Class Ii ◽  
In Vitro Differentiation ◽  
Rt Pcr ◽  
Hepatic Differentiation ◽  
Alizarin Red ◽  
Differentiation Potential

Mesenchymal stem cells (MSC) are isolated from bone marrow or other tissues, and have properties of self renewal and multilineage differentiation ability. The current study investigated the in vitro differentiation potential of porcine bone marrow derived MSCs into hepatocyte-like cells. The MSC were isolated from the bone marrow of adult miniature pigs (7 months old, T-type, PWG Micro-pig®, PWG Genetics, Seoul, Korea) and adherent cells with fibroblast-like morphology were cultured on plastic. Isolated MSCs were positive for CD29, CD44, CD73, CD90, and vimentin, and negative for CD34, CD45, major histocompatibility complex-class II (MHC-class II), and swine leukocyte antigen-DR (SLA-DR) by flow cytometry analysis. Further, trilineage differentiation of MSC into osteocytes (alkaline phosphatase, von Kossa and Alizarin red), adipocytes (Oil Red O), and chondrocytes (Alcian blue) was confirmed. Differentiation of MSC into hepatocyte-like cells was induced with sequential supplementation of growth factors, cytokines, and hormones for 21 days as described previously (Taléns-Visconti et al. 2006 World J. Gastroenterol. 12, 5834–5845). Morphological analysis, expression of liver-specific markers, and functional assays were performed to evaluate the hepatic differentiation of MSC. Under hepatogenic conditions, MSC acquired cuboidal morphology with cytoplasmic granules. These hepatocyte-like cells expressed α-fetoprotein (AFP), albumin (ALB), cytokeratin 18 (CK18), cytochrome P450 7A1 (CYP7A1), and hepatocyte nuclear factor 1 (HNF-1) markers by immunofluorescence assay. In addition, the expression of selected markers was demonstrated by Western blotting analysis. In accordance with these features, RT-PCR revealed transcripts of AFP, ALB, CK18, CYP7A1, and HNF-1α. Further, the relative expression levels of these transcripts were analysed by quantitative RT-PCR after normalizing to the expression of the endogenous control, glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Data were analysed statistically by one-way ANOVA using PASW statistics 18 (SPSS Inc., Chicago, IL, USA), and significance was considered at P < 0.05. The results showed that the relative expressions of selected marker genes in hepatocyte-like cells were significantly increased compared with that in untreated MSC. The generated hepatocyte-like cells showed glycogen storage as analysed by periodic acid-Schiff (PAS) staining. Moreover, the induced cells produced urea at Day 21 of culture compared with control MSC. In conclusion, our results indicate the potential of porcine MSC to differentiate in vitro into hepatocyte-like cells. Further studies on the functional properties of hepatocyte-like cells are needed to use porcine MSC as an ideal source for liver cell therapy and preclinical drug evaluation. This work was supported by Basic Science Research Program through the National Research Foundation (NRF), funded by the Ministry of Education, Science and Technology (2010-0010528) and the Next-Generation BioGreen 21 Program (No. PJ009021), Rural Development Administration, Republic of Korea.

scholarly journals Generation of in vivo neural stem cells using partially reprogrammed cells defective in in vitro differentiation potential

Oncotarget ◽  
2017 ◽  
Vol 8 (10) ◽  
pp. 16456-16462 ◽  
Author(s):  
Jong Soo Kim ◽  
Yean Ju Hong ◽  
Hyun Woo Choi ◽  
Hyuk Song ◽  
Sung June Byun ◽  
...  
Keyword(s):  
Stem Cells ◽  
Neural Stem Cells ◽  
In Vitro Differentiation ◽  
Differentiation Potential

Cell interactions and endoderm differentiation in cultured mouse embryos

Development ◽  
1981 ◽  
Vol 62 (1) ◽  
pp. 379-394
Author(s):  
Brigid L. M. Hogan ◽  
Rita Tilly
Keyword(s):  
Basement Membrane ◽  
Mouse Embryo ◽  
Cell Lineage ◽  
Giant Cells ◽  
Visceral Endoderm ◽  
Extraembryonic Endoderm ◽  
Type Iv ◽  
The Matrix ◽  
Parietal Endoderm

Morphological and biochemical evidence is presented that the visceral extraembryonic endoderm of the 6·5-day mouse embryo will differentiate into parietal endoderm when cultured in contact with extraembryonic ectoderm undergoing transition into trophoblast giant cells. Egg cylinders from 6·5-day embryos were dissected into embryonic and extraembryonic halves and cultured in suspension in vitro for up to 7 days. After 4 days, the endoderm cells of the extraembryonic fragments morphologically resemble parietal endoderm, are associated with a thick basement membrane and synthesize large amounts of the matrix proteins laminin and Type IV procollagen. A similar transition in phenotype is not seen in the endoderm of embryonic fragments, nor in visceral extraembryonic endoderm cells cultured in isolation. In another series of experiments, complete egg cylinders were dissected free of visceral endoderm ovei lying the extraembryonic ectoderm and then cultured in vitro. The visceral endoderm cells which recolonize the surface of the extraembryonic ectoderm develop a parietal endoderm phenotype and lay down a thick basement membrane. These results suggest that the differentiation of the extraembryonic endoderm of the early mouse embryo into visceral and parietal phenotypes can be influenced by local cell—cell or cell—substrate interactions, and is not determined solely by cell lineage.

Chicken embryonic brain: an in vivo model for verifying neural stem cell potency

2013 ◽  
Vol 119 (2) ◽  
pp. 512-519 ◽  
Author(s):  
Alex Kharazi ◽  
Michael L. Levy ◽  
Maria Cristina Visperas ◽  
Chih-Min Lin
Keyword(s):  
Stem Cell ◽  
Fetal Brain ◽  
Stem Cell Marker ◽  
In Vivo Model ◽  
Embryonic Brain ◽  
In Vitro Differentiation ◽  
Differentiation Potential ◽  
Marker Expression

Object The multipotency of neural stem cells (NSCs) can be assessed in vitro by detection of stage-specific markers in response to a suitable differentiation signal. This test is frequently used because it is fast and affordable. However, it is not clear how the in vitro potential for multilineage differentiation and stem cell marker expression would reflect the ability of NSCs to engraft into the brain following transplantation. The authors undertook this study to directly compare the in vitro potency and in vivo migration of human NSCs (hNSCs) expanded under conditions of gradually increased concentration of fetal bovine serum (FBS) as a maturation factor. Methods Human NSCs isolated from fetal brain were propagated in serum free media (SF-hNSCs) and in media containing 0.1% and 0.2% serum. At Passage 4 in tissue culture the NSCs were harvested and either differentiated in vitro or transplanted into the lateral ventricle of chicken embryonic brain at the late stage of its development (Hamburger and Hamilton Stage 26). The in vitro differentiation was evaluated by immunostaining with neural or glial specific markers, and the in vivo migration was assessed using immunohistology. Results The authors found that SF-hNSCs successfully engrafted into the chicken embryonic brain, which correlated with their ability to differentiate in vitro. NSCs grown at as low as 0.1% concentration of FBS failed to demonstrate the robust in vivo migration pattern but still preserved the capability to differentiate in vitro. Furthermore, NSCs generated in media containing a higher concentration of FBS (0.2%) lost both the in vivo engraftment and in vitro differentiation potential. Conclusions The present study suggests that marker expression and in vitro differentiation assays might not provide adequate information regarding the behavior of NSCs following their transplantation. The in vivo migration following injection into chicken embryonic brain may provide an important assay of the potency of NSCs.

Bone marrow CD34+/B220+ progenitors target the inflamed brain and display in vitro differentiation potential toward microglia

2006 ◽  
Vol 20 (12) ◽  
pp. 2081-2092 ◽  
Author(s):  
N. Davoust ◽  
C. Vuaillat ◽  
G. Cavillon ◽  
C. Domenget ◽  
E. Hatterer ◽  
...  
Keyword(s):  
Bone Marrow ◽  
In Vitro Differentiation ◽  
Differentiation Potential

In vitro differentiation potential of a new human osteosarcoma cell line (HOS 58)

1998 ◽  
Vol 63 (2) ◽  
pp. 81-91 ◽  
Author(s):  
Heide Siggelkow ◽  
Christopher Niedhart ◽  
Wiebke Kurre ◽  
Angela Ihbe ◽  
Andreas Schulz ◽  
...  
Keyword(s):  
Cell Line ◽  
Osteosarcoma Cell Line ◽  
Osteosarcoma Cell ◽  
In Vitro Differentiation ◽  
Human Osteosarcoma Cell Line ◽  
Differentiation Potential ◽  
Human Osteosarcoma ◽  
Human Osteosarcoma Cell

In vitro differentiation potential of human haematopoietic CD34+cells towards pancreatic β-cells

2016 ◽  
Vol 40 (10) ◽  
pp. 1084-1093 ◽  
Author(s):  
Manne Mudhu Sunitha ◽  
Lokanathan Srikanth ◽  
Pasupuleti Santhosh Kumar ◽  
Chodimella Chandrasekhar ◽  
Potukuchi Venkata Gurunadha Krishna Sarma
Keyword(s):  
In Vitro Differentiation ◽  
Β Cells ◽  
Pancreatic Β Cells ◽  
Differentiation Potential ◽  
Cd34 Cells
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