The effects of human umbilical cord perivascular cells on rat hepatocyte structure and functional polarity

2013 ◽  
Vol 91 (3) ◽  
pp. 140-147 ◽  
Author(s):  
Alejandro Gómez-Aristizábal ◽  
John Edward Davies
Keyword(s):  
Umbilical Cord ◽  
Ex Vivo ◽  
Human Umbilical Cord ◽  
Perivascular Cells ◽  
Transmission Electron ◽  
External Organization ◽  
Bioartificial Livers ◽  
Functional Polarization ◽  
Hepatocyte Growth

Hepatocyte culture is a useful tool for the study of their biology and the development of bioartificial livers. However, many challenges have to be overcome since hepatocytes rapidly lose their normal phenotype in vitro. We have recently demonstrated that human umbilical cord perivascular cells (HUCPVCs) are able to provide support to hepatocytes. In the present study we go further into exploring the effects that HUCPVCs have in the functional polarization, and both the internal and external organization, of hepatocytes. Also, we investigate HUCPVC–hepatocyte crosstalk by tracking both the effects of HUCPVCs on hepatocyte transcription factors and those of hepatocytes on the expression of hepatotrophic factors in HUCPVCs. Our results show that HUCPVCs maintain the functional polarity of hepatocytes ex vivo, as judged by the secretion of fluorescein into bile canaliculi, for at least 40 days. Transmission electron microscopy revealed that hepatocytes in coculture organize in an organoid-like structure embedded in extracellular matrix surrounded by HUCPVCs. In coculture, hepatocytes displayed a higher expression of C/EBPα, implicated in maintenance of the mature hepatocyte phenotype, and HUCPVCs upregulated hepatocyte growth factor and Jagged1 indicating that these genes may play important roles in HUCPVC–hepatocyte interactions.

Abstract 1144: Successful Surface-Aminated Nanofiber Expansion of Human Umbilical Cord-Derived CD133+ Cells Leads to Augmentation of Angiogenic Functionality In Vitro and In Vivo

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Hiranmoy Das ◽  
Matthew Joseph ◽  
Nasreen Abdulhameed ◽  
Hai-Quan Mao ◽  
Vincent J Pompili
Keyword(s):  
Umbilical Cord ◽  
Hind Limb ◽  
Ex Vivo ◽  
Flow Cytometric Analysis ◽  
Surface Expression ◽  
Capillary Density ◽  
Therapeutic Angiogenesis ◽  
Human Umbilical Cord

Background: Umbilical cord blood (UCB) and marrow-derived CD133+ cells have been shown to mediate encouraging effects on therapeutic angiogenesis in both animal models and early clinical trials. Low numbers of CD133+ cells derived from a single donor have been a limitation of use of these cells in cardiovascular therapy. We hypothesized that an ex vivo aminated nanofiber system combined with cytokine supplementation would provide optimized topographical and biochemical signals to allow the expansion and potential functional augmentation of CD133+ cells without promoting terminal differentiation. Methods and Results: Human UCB derived CD133+ progenitor cells were isolated by MACS sorting and ex vivo expanded on aminated nanofiber plates with cytokine rich media. Cells harvested 10 days after expansion demonstrated a 225X increase in total number. Flow cytometric analysis demonstrated CD133–24%, CD34–93%, CXCR4–97%, LFA-97% surface expression. The expanded cells can uptake AcLDL efficiently and demonstrate a 2.3X increase in transwell migration to SDF-1 as compared to fresh UCB CD133+ cells. In vitro analysis revealed that expanded cells have potential to differentiate into endothelial or smooth muscle phenotype as demonstrated with CD31, vWF, VCAM-1 and F-pholloidin, α-actin, and SM myosin heavy chain immunocytochemistry when re-cultured for 14d in EGM2 or SMBM respectively. RT-QPCR analysis of 1% O 2 exposed (hypoxic) cells demonstrated a 2X increase in VEGF and 3X increase in IL-8 gene expression compared to normoxic control. In vivo functionality in a NOD/SCID mouse hind limb ischemic model demonstrated that mice treated with 5 x 10 6 expanded cells (n=7) augmented blood flow ratio (ischemic/control limb) as compared to mice treated with CD133+ cells (n=7) and control (n=7) at 28d. (control 0.32±.02 vs. UCB133+ 0.37±.02 vs. expanded cells 0.50±.04 p<0.01) Capillary density in ischemic hind-limb was increased at 28d (control 62.5±5.4 vs. expanded cell 97.6±2.5 p< 0.01) Conclusions: These studies demonstrate successful high level expansion of UCB derived CD133+ cells into functionally potent stem cells which have the capacity to differentiate into vascular cells and promote in vivo neovascularization.

scholarly journals In VitroDifferentiation of First Trimester Human Umbilical Cord Perivascular Cells into Contracting Cardiomyocyte-Like Cells

2016 ◽  
Vol 2016 ◽  
pp. 1-13 ◽  
Author(s):  
Peter Szaraz ◽  
Matthew Librach ◽  
Leila Maghen ◽  
Farwah Iqbal ◽  
Tanya A. Barretto ◽  
...  
Keyword(s):  
Umbilical Cord ◽  
Cardiac Regeneration ◽  
First Trimester ◽  
Human Umbilical Cord ◽  
Perivascular Cells ◽  
Differentiation Potential ◽  
Mortality And Morbidity ◽  
Cardiomyogenic Differentiation

Myocardial infarction (MI) causes an extensive loss of heart muscle cells and leads to congestive heart disease (CAD), the leading cause of mortality and morbidity worldwide. Mesenchymal stromal cell- (MSC-) based cell therapy is a promising option to replace invasive interventions. However the optimal cell type providing significant cardiac regeneration after MI is yet to be found. The aim of our study was to investigate the cardiomyogenic differentiation potential of first trimester human umbilical cord perivascular cells (FTM HUCPVCs), a novel, young source of immunoprivileged mesenchymal stromal cells. Based on the expression of cardiomyocyte markers (cTnT, MYH6, SIRPA, and CX43) FTM and term HUCPVCs achieved significantly increased cardiomyogenic differentiation compared to bone marrow MSCs, while their immunogenicity remained significantly lower as indicated by HLA-A and HLA-G expression and susceptibility to T cell mediated cytotoxicity. When applying aggregate-based differentiation, FTM HUCPVCs showed increased aggregate formation potential and generated contracting cells within 1 week of coculture, making them the first MSC type with this ability. Our results indicate that young FTM HUCPVCs have superior cardiomyogenic potential coupled with beneficial immunogenic properties when compared to MSCs of older tissue sources, suggesting thatin vitropredifferentiation could be a potential strategy to increase their effectivenessin vivo.

Human umbilical cord perivascular cells (HUC-PVCs) protect axons from degeneration in an in vitro peripheral nervous system injury model

Cytotherapy ◽  
2015 ◽  
Vol 17 (6) ◽  
pp. S56
Author(s):  
Tanya Barretto ◽  
Katya Park ◽  
Leila Maghen ◽  
Shlomit Kenigsberg ◽  
Andree Gauthier-Fisher ◽  
...  
Keyword(s):  
Nervous System ◽  
Peripheral Nervous System ◽  
Umbilical Cord ◽  
Human Umbilical Cord ◽  
Perivascular Cells ◽  
Injury Model

Assessment of the angiogenic potential of human umbilical cord-derived perivascular cells and bone marrow MSCs using novel functional in vitro assays

Cytotherapy ◽  
2015 ◽  
Vol 17 (6) ◽  
pp. S54-S55
Author(s):  
Farwah Iqbal ◽  
Peter Szaraz ◽  
Matthew Librach ◽  
Leila Maghen ◽  
Tanya Barretto ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Umbilical Cord ◽  
In Vitro Assays ◽  
Human Umbilical Cord ◽  
Bone Marrow Mscs ◽  
Perivascular Cells ◽  
Angiogenic Potential

Optimization of xeno-free expansion conditions for human umbilical cord-derived perivascular cells using functional immunophenotyping and in vitro potency assays

Cytotherapy ◽  
2015 ◽  
Vol 17 (6) ◽  
pp. S82-S83
Author(s):  
Matthew Librach ◽  
Schreiber Pereira ◽  
Leila Maghen ◽  
Farwah Iqbal ◽  
Tanya Barretto ◽  
...  
Keyword(s):  
Umbilical Cord ◽  
Human Umbilical Cord ◽  
Perivascular Cells ◽  
Free Expansion

scholarly journals Human umbilical perivascular cells (HUCPVCs): a novel source of mesenchymal stromal-like (MSC) cells to support the regeneration of the testicular niche

Reproduction ◽  
2017 ◽  
Vol 153 (1) ◽  
pp. 85-95 ◽  
Author(s):  
Leila Maghen ◽  
Ekaterina Shlush ◽  
Itai Gat ◽  
Melissa Filice ◽  
Tanya A Barretto ◽  
...  
Keyword(s):  
Germ Cell ◽  
Ex Vivo ◽  
Somatic Cells ◽  
Human Umbilical Cord ◽  
Sequencing Analysis ◽  
Feeder Cells ◽  
Perivascular Cells ◽  
Male Factor

The expansion of functional testicular biopsy-derived human spermatogonial stem cells (hSSC)ex vivomay enable the restoration of fertility in pre-pubertal males having undergone gonadotoxic therapies or men with severe male factor infertility. Various somatic cells are known to regulate SSC homeostasis and spermatogenesis in the developing and adult testis. Prior attempts to recapitulate this niche demonstrated the requirement of feeder cells, such as endogenous testicular somatic cells, for germ cell expansionex vivo. However, this strategy has limitations in the expansion of hSSCs from tissue biopsies for which spermatogenesis is absent or defective. Our aim was to evaluate the first trimester human umbilical cord perivascular cells (FTM HUCPVCs), a novel source of mesenchymal stromal-like cells (MSCs), as potential human feeder cells for standardized hSSC expansionex vivo.Targeted RNA sequencing analysis demonstrated that CD90+ve FTM HUCPVCs expandedin vitrounder germ cell culture conditions express a profile of targeted testicular-associated transcripts that is similar to cultured human CD90+ve testicular adherent cells (hTACs) and secrete LIF, FGF2 and BMP4, key growth factors known to regulate spermatogenesis. We also demonstrated that mitotically inactivated FTM HUCPVCs support the expansion of mouse germ cells and putative SSCsex vivoand that FTM HUCPVC transplantation promotesin vivogerm cell regeneration after mono-2-ethylhexyl phthalate (MEHP)-induced seminiferous tubule damage in a murine model, including a partial reconstitution of tubular cellular architecture and reestablishment of DAZL and acrosin+ve germ cell layers. Together, these data suggest that FTM HUCPVCs have phenotypical and functional properties that may support repair of the human testicular niche.

scholarly journals In vitro generation of Sertoli-like and haploid spermatid-like cells from human umbilical cord perivascular cells

2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Ekaterina Shlush ◽  
Leila Maghen ◽  
Sonja Swanson ◽  
Shlomit Kenigsberg ◽  
Sergey Moskovtsev ◽  
...  
Keyword(s):  
Umbilical Cord ◽  
Human Umbilical Cord ◽  
Perivascular Cells

In vitro immunologic properties of human umbilical cord perivascular cells

Cytotherapy ◽  
2008 ◽  
Vol 10 (2) ◽  
pp. 174-181 ◽  
Author(s):  
J. Ennis ◽  
C. Götherström ◽  
K. Le Blanc ◽  
J.E. Davies
Keyword(s):  
Umbilical Cord ◽  
Human Umbilical Cord ◽  
Perivascular Cells ◽  
Immunologic Properties

scholarly journals Charachterization of umbilical cord mesenchymal stromal cells during long-term expansion in vitro

2018 ◽  
Vol 10 (1) ◽  
pp. 11-19
Author(s):  
A. A. Aizenshtadt ◽  
M. A. Skazina ◽  
E. A. Kotelevskaya ◽  
L. V. Yelsukova ◽  
T. L. Zolina ◽  
...  
Keyword(s):  
Umbilical Cord ◽  
Stromal Cells ◽  
Ex Vivo ◽  
Normal Karyotype ◽  
Telomerase Activity ◽  
Human Umbilical Cord ◽  
Critical Question ◽  
Karyotype Morphology

One of the clinicians’ major concerns is the biological safety of MSC. The critical question for clinical application of human MSC is their ability to undergo spontaneous malignant transformation in a recipient organism. The goal of our research was to study umbilical cord hMSC proliferative and differentiation capacities, karyotype stability, telomerase activity and telomere length, oncomarkers expression and tumorigenicity during long-term (6 months) cultivation ex vivo. Here we report on the establishing the primary culture of human umbilical cord MSC, MSC_0714, that was capable to proliferate ex vivo for up to 59 passages (6 months). During this period, the cells preserved their normal karyotype, morphology and MSC immunophenotype. Telomeres started to shorten only after the passage 20, while hTERT was inactive in these cells for the whole period of expansion. At the beginning of cultivation the number of SA-β-gal positive cells did not exceeded 3-5%, after the 22th passage their number started to increase and reached 49% at the passage 57. Thus, it was shown that MSC during long-term culture retain their characteristics and undergo cell senescence.

Sign in / Sign up

Export Citation Format

Share Document