chromosome-preparation-from-cultured-cells

Chromosome Preparation From Cultured Cells

Chromosome (cytogenetic) analysis is widely used for the detection of chromosome instability. When followed by G-banding and molecular techniques such as ...

The isolation and characterization of embryonic and adult stem cells from higher-order mammalian species will enhance the understanding of the biology and therapeutic application of stem cells. The aim of this study was to purify rhesus mesenchymal stem cells (MSCs) from adult bone marrow and to characterize functionally their abilities to differentiate along diverse lineages. Adherent cells from adult rhesus macaque bone marrow were characterized for their growth characteristics, lineage differentiation, cell-surface antigen expression, telomere length, chromosome content, and transcription factor gene expression. Rhesus bone marrow MSCs (BMSCs) are very heterogeneous, composed of primarily long, thin cells and some smaller, round cells. The cells are capable of differentiating along osteogenic, chondrogenic, and adipogenic lineages in vitro. The cell morphology and multipotential differentiation capabilities are maintained throughout extended culture. They express CD59, CD90 (Thy-1), CD105, and HLA-1 and were negative for hematopoietic markers such as CD3, CD4, CD8, CD11b, CD13, CD34, and platelet endothelial cell adhesion molecule-1 (CD31). BMSCs were also demonstrated to express the mRNA for important stem cell-related transcription factors such as Oct-4, Sox-2, Rex-1, and Nanog. Rhesus BMSCs have a normal chromosome content, and the shortening of telomeres is minimal during early passages. These data demonstrate that BMSCs isolated from rhesus macaques have a high degree of commonality with MSCs isolated from other species. Therefore, isolation of these cells provides an effective and convenient method for rapid expansion of pluripotent rhesus MSCs.

Characterization of multipotent mesenchymal stem cells from the bone marrow of rhesus macaques.

Charakterisierung von multipotenten mesenchymalen Stammzellen aus dem Knochenmark von Rhesusaffen.

We report on the phenotypic features of a patient with partial trisomy of the long arm of chromosome 16 due to an unbalanced Y;16 translocation (46,X,der[Y]t[Y;16] [q12;q22]pat). The patient was noted to have craniofacial anomalies and developmental delay, but no other major malformations. The father, a balanced Y;16 translocation carrier, has apparently normal fertility.

Phenotypic features of a boy with trisomy of 16q22-->qter due to paternal Y; 16 translocation.

Mesenchymal stem cells (MSC) derived from bone marrow stem cells (BMSC) and adipose tissue stem cells (ASC) of humans and rhesus macaques were evaluated for their cell cycle properties during protracted culture in vitro. Human ASCs (hASC) and rhesus BMSCs (rBMSC) underwent significantly more total population doublings than human BMSCs (hBMSC) and rhesus ASCs (rASC). The cell cycle profile of all MSCs was altered as cultures aged. hMSCs underwent an increase in the frequency of cells in the S phase at P20 and P30. However, rhesus MSCs from both sources developed a distinct polyploid population of cells at P20, which progressed to aneuploidy by P30. Karyotype analysis of MSCs revealed the development of tetraploid or aneuploid karyotypes in the rhesus cells at P20 or P30. Analysis of the transcriptome of the MSCs from early and late passages revealed significant alterations in the patterns of gene expression (8.8% of the genes were differentially expressed in hBMSCs versus hASCs, and 5.5% in rBMSCs versus rASCs). Gene expression changes were much less evident within the same cell type as aging occurred (0.7% in hMSCs and 0.9% in rMSC). Gene ontology analysis showed that functions involved in protein catabolism and regulation of pol II transcription were overrepresented in rASCs, whereas the regulation of I kappa B/nuclear factor-kappaB cascade were overrepresented in hBMSCs. Functional analysis of genes that were differentially expressed in rASCs and hBMSCs revealed that pathways involved in cell cycle, cell cycle checkpoints, protein-ubiquitination, and apoptosis were altered.

Long-term in vitro expansion alters the biology of adult mesenchymal stem cells.

Mesenchymaler Stammzellen (MSC) abgeleitet von Knochenmark-Stammzellen (BMSC) und Fettgewebe Stammzellen (ASC) des Menschen und Rhesus-Makaken wurden f&uuml;r ihre Zellzyklus-Eigenschaften w&auml;hrend langwierige Kultur in-vitro-ausgewertet. Menschliche ASCs (hASC) und Rhesus-BMSCs (rBMSC) wurde deutlich mehr Gesamtbev&ouml;lkerung Verdoppelungen als menschliche BMSCs (hBMSC) und Rhesus ASCs (rASC). Das Zellzyklus Profil alle MSCs &auml;nderte sich wie im Alter Kulturen. hMSCs wurde ein Anstieg der H&auml;ufigkeit von Zellen in der S-Phase bei P20 und P30. Rhesus MSCs aus beiden Quellen entwickelt jedoch eine unterschiedliche polyploide Bev&ouml;lkerung von Zellen bei P20, die zu Aneuploidie von P30 Motiven. Karyotyp Analyse der MSCs ergab die Entwicklung von tetraploide oder aneuploider Karyotyp der Rhesus-Zellen bei P20 oder P30. Analyse der das Transkriptom des MSCs aus fr&uuml;hen und sp&auml;ten Durchg&auml;nge ergab wesentliche Ver&auml;nderungen im Muster Genexpression (8,8 % der Gene wurden differentiell hBMSCs versus hASCs und 5,5 % in rBMSCs gegen rASCs ausgedr&uuml;ckt). Gen Ausdruck &Auml;nderungen waren viel weniger offensichtlich innerhalb des gleichen Typs der Zelle, wie Alterung (0,7 % in hMSCs und 0,9 % in rMSC) aufgetreten ist. Gen Ontologie Analyse zeigte, dass Funktionen Protein Katabolismus und Regulierung der Pol II Transkription beteiligt &uuml;berrepr&auml;sentiert in rASCs waren, w&auml;hrend die Verordnung ich Kappa B/nuclear Factor-KappaB Kaskade in hBMSCs &uuml;berrepr&auml;sentiert waren. Funktionelle Analyse von Genen, die differentiell in rASCs und hBMSCs ausgedr&uuml;ckt wurden ergab, dass Wege Zellzyklus, Zellzyklus Checkpoints, Protein-Ubiquitination und Apoptose beteiligt ver&auml;ndert wurden.

Article	Year
Characterization of multipotent mesenchymal stem cells from the bone marrow of rhesus macaques. Stem cells and development\| PubMed ID: 16137233	2005
Phenotypic features of a boy with trisomy of 16q22-->qter due to paternal Y; 16 translocation. Clinical dysmorphology\| PubMed ID: 16155418	2005
Long-term in vitro expansion alters the biology of adult mesenchymal stem cells. Cancer research\| PubMed ID: 18519682	2008

Fern Tsien

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