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Nanoparticle labeling identifies slow cycling human endometrial stromal cells.

Xiang L, Chan RW, Ng EH, Yeung WS - Stem Cell Res Ther (2014)

Bottom Line: It remains unclear whether slow-cycling cells exist in the human endometrium.They also differentiate into multiple mesenchymal lineages and the expression of lineage specific markers was lower than that of non-FPC.In summary, nanoparticle labeling is a promising tool for the identification of putative somatic stem or progenitor cells when their surface markers are undefined.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Evidence suggests that the human endometrium contains stem or progenitor cells that are responsible for its remarkable regenerative capability. A common property of somatic stem cells is their quiescent state. It remains unclear whether slow-cycling cells exist in the human endometrium. We hypothesized that the human endometrium contains a subset of slow-cycling cells with somatic stem cell properties. Here, we established an in vitro stem cell assay to isolate human endometrial-derived mesenchymal stem-like cells (eMSC).

Methods: Single-cell stromal cultures were initially labeled with fluorescent nanoparticles and a small population of fluorescent persistent cells (FPC) remained after culture of 21 days. Two populations of stromal cells, namely FPC and non-FPC were sorted.

Results: Quantitative analysis of functional assays demonstrated that the FPC had higher colony forming ability, underwent more rounds of self-renewal and had greater enrichment of phenotypically defined prospective eMSC markers: CD146+/CD140b+ and W5C5+ than the non-FPC. They also differentiate into multiple mesenchymal lineages and the expression of lineage specific markers was lower than that of non-FPC. The FPC exhibit low proliferation activities. A proliferation dynamics study revealed that more FPC had a prolonged G1 phase.

Conclusions: With this study we present an efficient method to label and isolate slow-proliferating cells obtained from human endometrial stromal cultures without genetic modifications. The FPC population could be easily maintained in vitro and are of interest for tissue-repair and engineering perspectives. In summary, nanoparticle labeling is a promising tool for the identification of putative somatic stem or progenitor cells when their surface markers are undefined.

No MeSH data available.


Related in: MedlinePlus

Expression of pluripotent and self-renewal genes. The relative gene expression levels of (A)NANOG, (B)SOX2, (C)OCT-4 and (D)BMI-1 of passage 1 and 2 (P1, P2) FPC (white bars) and non-FPC (black bars) in comparison to human embryonic stem cells (hESC, grey bars) as positive control. Results are reported as means ± SEM, n = 3. FPC, fluorescent persistent cells; SEM, standard error of the mean.
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Figure 6: Expression of pluripotent and self-renewal genes. The relative gene expression levels of (A)NANOG, (B)SOX2, (C)OCT-4 and (D)BMI-1 of passage 1 and 2 (P1, P2) FPC (white bars) and non-FPC (black bars) in comparison to human embryonic stem cells (hESC, grey bars) as positive control. Results are reported as means ± SEM, n = 3. FPC, fluorescent persistent cells; SEM, standard error of the mean.

Mentions: To gain further insight into the molecular properties of the two functionally distinct populations, we performed real-time PCR to determine their expression of genes associated with pluripotency in human embryonic stem cells (NANOG, SOX2, OCT4) [25], and self-renewal in somatic stem cells (BMI-1) [26]. Comparison of the FPC and non-FPC transcript expression at P1 revealed no differences for these genes (Figure 6). Interestingly, an up-regulation trend for the four genes (Figure 6A-D, n = 3) was observed in cells of the secondary FPC when compared to that of primary FPC and non-FPC, though the difference had not yet reached statistical significance (P = 0.10) probably because of the small sample size.


Nanoparticle labeling identifies slow cycling human endometrial stromal cells.

Xiang L, Chan RW, Ng EH, Yeung WS - Stem Cell Res Ther (2014)

Expression of pluripotent and self-renewal genes. The relative gene expression levels of (A)NANOG, (B)SOX2, (C)OCT-4 and (D)BMI-1 of passage 1 and 2 (P1, P2) FPC (white bars) and non-FPC (black bars) in comparison to human embryonic stem cells (hESC, grey bars) as positive control. Results are reported as means ± SEM, n = 3. FPC, fluorescent persistent cells; SEM, standard error of the mean.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4230801&req=5

Figure 6: Expression of pluripotent and self-renewal genes. The relative gene expression levels of (A)NANOG, (B)SOX2, (C)OCT-4 and (D)BMI-1 of passage 1 and 2 (P1, P2) FPC (white bars) and non-FPC (black bars) in comparison to human embryonic stem cells (hESC, grey bars) as positive control. Results are reported as means ± SEM, n = 3. FPC, fluorescent persistent cells; SEM, standard error of the mean.
Mentions: To gain further insight into the molecular properties of the two functionally distinct populations, we performed real-time PCR to determine their expression of genes associated with pluripotency in human embryonic stem cells (NANOG, SOX2, OCT4) [25], and self-renewal in somatic stem cells (BMI-1) [26]. Comparison of the FPC and non-FPC transcript expression at P1 revealed no differences for these genes (Figure 6). Interestingly, an up-regulation trend for the four genes (Figure 6A-D, n = 3) was observed in cells of the secondary FPC when compared to that of primary FPC and non-FPC, though the difference had not yet reached statistical significance (P = 0.10) probably because of the small sample size.

Bottom Line: It remains unclear whether slow-cycling cells exist in the human endometrium.They also differentiate into multiple mesenchymal lineages and the expression of lineage specific markers was lower than that of non-FPC.In summary, nanoparticle labeling is a promising tool for the identification of putative somatic stem or progenitor cells when their surface markers are undefined.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Evidence suggests that the human endometrium contains stem or progenitor cells that are responsible for its remarkable regenerative capability. A common property of somatic stem cells is their quiescent state. It remains unclear whether slow-cycling cells exist in the human endometrium. We hypothesized that the human endometrium contains a subset of slow-cycling cells with somatic stem cell properties. Here, we established an in vitro stem cell assay to isolate human endometrial-derived mesenchymal stem-like cells (eMSC).

Methods: Single-cell stromal cultures were initially labeled with fluorescent nanoparticles and a small population of fluorescent persistent cells (FPC) remained after culture of 21 days. Two populations of stromal cells, namely FPC and non-FPC were sorted.

Results: Quantitative analysis of functional assays demonstrated that the FPC had higher colony forming ability, underwent more rounds of self-renewal and had greater enrichment of phenotypically defined prospective eMSC markers: CD146+/CD140b+ and W5C5+ than the non-FPC. They also differentiate into multiple mesenchymal lineages and the expression of lineage specific markers was lower than that of non-FPC. The FPC exhibit low proliferation activities. A proliferation dynamics study revealed that more FPC had a prolonged G1 phase.

Conclusions: With this study we present an efficient method to label and isolate slow-proliferating cells obtained from human endometrial stromal cultures without genetic modifications. The FPC population could be easily maintained in vitro and are of interest for tissue-repair and engineering perspectives. In summary, nanoparticle labeling is a promising tool for the identification of putative somatic stem or progenitor cells when their surface markers are undefined.

No MeSH data available.


Related in: MedlinePlus