Ultrastructural and immunohistochemical studies of Wharton's jelly umbilical cord cells

Takechi, Kimihiro; Kuwabara, Yoshinori; Mizuno, Masahiko

doi:10.1016/s0143-4004(05)80264-4

Cited by 124 publications

(104 citation statements)

References 14 publications

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“…Thus, immunohistochemistry demonstrated the presence of three specific cytoskeletal markers-α-actin, desmin, and vimentin-which correlates with the in situ characterization of WJ cells by Takechi et al [40], Kobayashi et al [44], and Kadner et al [37,38]. Furthermore, due to their reactivity with the 3G5 monoclonal antibody [45], HUCPV cells appear to be similar to another perivascular mesenchymal precursor, the pericyte [46][47][48][49].…”

Section: Discussionmentioning

confidence: 67%

Human Umbilical Cord Perivascular (HUCPV) Cells: A Source of Mesenchymal Progenitors

et al. 2005

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We describe the isolation of a nonhematopoietic (CD45 -, CD34 -, SH2 + , SH3 + , Thy-1 + , CD44 + ) human umbilical cord perivascular (HUCPV) cell population. Each HUCPV cell harvest (2-5 × 10 6 , depending on the length of cord available) gave rise to a morphologically homogeneous fibroblastic cell population, which expressed α-actin, desmin, vimentin, and 3G5 (a pericyte marker) in culture. We determined the colony-forming unit-fibroblast (CFU-F) frequency of primary HUCPV cells to be 1:333 and the doubling time, which was 60 hours at passage 0 (P0), decreased to 20 hours at P2. This resulted in a significant cell expansion, producing over 10 10 HUCPV cells within 30 days of culture. Furthermore, HUCPV cells cultured in nonosteogenic conditions contained a subpopulation that exhibited a functional osteogenic phenotype and elaborated bone nodules. The frequency of this CFU-osteogenic subpopulation at P1 was 2.6/10 5 CFU-F, which increased to 7.5/10 5 CFU-F at P2. Addition of osteogenic supplements to the culture medium resulted in these frequencies increasing to 1.2/10 4 and 1.3/10 4 CFU-F, respectively, for P1 and P2. CFU-O were not seen at P0 in either osteogenic or non-osteogenic culture conditions, but P0 HUCPV cells did contain a 20% subpopulation that presented neither class I nor class II cell-surface major histocompatibility complexes (MHC -/-). This population increased to 95% following passage and cryopreservation (P5). We conclude that, due to their rapid doubling time, high frequencies of CFU-F and CFU-O, and high MHC -/-phenotype, HUCPV cells represent a significant source of cells for allogeneic mesenchymal cell-based therapies. Stem Cells 2005;23:220-229

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Section: Discussionmentioning

confidence: 67%

Human Umbilical Cord Perivascular (HUCPV) Cells: A Source of Mesenchymal Progenitors

et al. 2005

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“…It also has angiogenic and metabolic roles for the umbilical circulation. Since 1990, stromal cells, which basically resemble mesenchymal fibroblasts found elsewhere during in utero development, were identified in human WJ (Takechi et al 1993, Nanaev et al 1997. Stemness of these cells has been demonstrated by in vitro differentiation (Lu et al 2006) and identification of MSCs membrane-specific markers.…”

Section: Introductionmentioning

confidence: 99%

Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton's jelly in the horse

Iacono

Brunori²,

Pirrone³

et al. 2012

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Mesenchymal stem cells (MSCs) have been derived from multiple sources of the horse including umbilical cord blood (UCB) and amnion. This work aimed to identify and characterize stem cells from equine amniotic fluid (AF), CB and Wharton's Jelly (WJ). Samples were obtained from 13 mares at labour. AF and CB cells were isolated by centrifugation, while WJ was prepared by incubating with an enzymatic solution for 2 h. All cell lines were cultured in DMEM/TCM199 plus fetal bovine serum. Fibroblast-like cells were observed in 7/10 (70%) AF, 6/8 (75%) CB and 8/12 (66.7%) WJ samples. Statistically significant differences were found between cell-doubling times (DTs): cells isolated from WJ expanded more rapidly (2.0G0.6 days) than those isolated from CB (2.6G1.3 days) and AF (2.3G1.0 days) (P!0.05). Positive von Kossa and Alizarin Red S staining confirmed osteogenesis. Alcian Blue staining of matrix glycosaminoglycans illustrated chondrogenesis and positive Oil Red O lipid droplets staining suggested adipogenesis. All cell lines isolated were positive for CD90, CD44, CD105; and negative for CD34, CD14 and CD45. These findings suggest that equine MSCs from AF, UCB and WJ appeared to be a readily obtainable and highly proliferative cell lines from a uninvasive source that may represent a good model system for stem cell biology and cellular therapy applications in horses. However, to assess their use as an allogenic cell source, further studies are needed for evaluating the expression of markers related to cell immunogenicity.

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“…Since the first isolation of MSCs from bone marrow (5), many different tissues were found to harbor MSCs as well, including adipose tissue (6,7), tendon tissue (8), umbilical cord blood (9), umbilical cord matrix (10,11), and peripheral blood (12,13). MSCs from all sources show a spindle shaped morphology but differ in growth characteristics, differentiation potential (14,15), and gene expression of chemokine receptors (16).…”

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confidence: 99%

Comparative immunophenotyping of equine multipotent mesenchymal stromal cells: An approach toward a standardized definition

et al. 2014

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Horses are an approved large animal model for therapies of the musculoskeletal system. Especially for tendon disease where cell-based therapy is commonly used in equine patients, the translation of achieved results to human medicine would be a great accomplishment. Immunophenotyping of equine mesenchymal stromal cells (MSCs) remains the last obstacle to meet the criteria of the International Society for Cellular Therapy (ISCT) definition of human MSCs. Therefore, the surface antigen expression of CD 29, CD 44, CD 73, CD 90, CD 105, CD 14, CD 34, CD 45, CD 79a, and MHC II in equine MSCs from adipose tissue, bone marrow, umbilical cord blood, umbilical cord tissue, and tendon tissue was analyzed using flow cytometry. Isolated cells from the different sources and donors varied in their expression pattern of MSC-defining antigens. In particular, CD 90 and 105 showed most heterogeneity. However, cells from all samples were robustly positive for CD 29 and CD 44, while being mostly negative for CD 73 and the exclusion markers CD 14, CD 34, CD 45, CD 79a and MHC II. Furthermore, it was evident that enzymes used for cell detachment after in vitro-culture affected the detection of antigen expression. These results emphasize the need of standardization of MSC isolation, culturing, and harvesting techniques. As the equine MSCs did not meet all criteria the ISCT defined for human MSCs, further investigations for a better characterization of the cell type should be conducted. V C 2014 International Society for Advancement of Cytometry

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Ultrastructural and immunohistochemical studies of Wharton's jelly umbilical cord cells

Cited by 124 publications

References 14 publications

Human Umbilical Cord Perivascular (HUCPV) Cells: A Source of Mesenchymal Progenitors

Human Umbilical Cord Perivascular (HUCPV) Cells: A Source of Mesenchymal Progenitors

Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton's jelly in the horse

Comparative immunophenotyping of equine multipotent mesenchymal stromal cells: An approach toward a standardized definition

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