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Updated: Apr 19, 2026

Isolation of CD146+ Resident Lung Mesenchymal Stromal Cells from Rat Lungs
Published on: June 17, 2016
[CD271 enrichment does not help isolating mesenchymal stromal cells from]
Abstract:
Reports on the isolation of mesencnymal stromal cells (MSCs) from granulocyte colony stimulating factor mobilized peripheral blood (G-CSF-mobilized PB) using regular culturing techniques are controversial. Enrichment techniques such as CD133 isolation have increased the success rates. CD271 is a well-known marker for enrichment of MSCs from bone marrow (BM). In the present study, we aimed to find out whether CD271 enrichment can help isolation of MSCs from G-CSF-mobilized PB. Five G-CSF-mobilized PB samples were collected from the remnant parts of the bags used for BM transplantation. Five BM samples were used as the control. Mononuclear cells (MNCs) from both resources were collected and underwent magnetic sorting for CD271-positive cells. The isolated cells were cultured, undergoing flowcytometry and differentiation assays to determine if they fulfill MSCs characteristics. CD271-positive portion of G-CSF-mobilized PB did not yield any cell outgrowth but the BM counterpart could successfully form MSC colonies. Although the percentage of CD271+ cells showed no difference between BM-MNCs and G-CSF-mobilized PB-MNCs, hematopoietic markers such as CD45, CD34 and CD133 composed a higher percentage of CD271-positive cells in the G-CSF-mobilized PB group. Results obtained indicated that CD271 enrichment does not help isolation of MSCs from G-CSF-mobilized PB. In this source, almost all of the CD271+ cells are from hematopoietic origin and the frequency of MSCs is so low that possibly during the process of cell isolation most of them are lost and the isolation fails.
Insights
CD271 enrichment does not improve mesenchymal stromal cell (MSC) isolation from G-CSF-mobilized peripheral blood. This method fails because CD271+ cells in mobilized blood are primarily hematopoietic, not MSCs.
Area of Science:
- Cell Biology
- Hematology
- Stem Cell Research
Background:
- Mesenchymal stromal cell (MSC) isolation from G-CSF-mobilized peripheral blood (PB) is challenging.
- Enrichment strategies, like CD133 isolation, have shown promise.
- CD271 is a known MSC marker in bone marrow (BM).
Purpose of the Study:
- To investigate if CD271 enrichment can enhance MSC isolation from G-CSF-mobilized PB.
- To compare CD271+ cell populations from G-CSF-mobilized PB and BM.
Main Methods:
- Collected G-CSF-mobilized PB and BM samples.
- Isolated CD271-positive mononuclear cells (MNCs) using magnetic sorting.
- Cultured isolated cells and performed flow cytometry and differentiation assays.
Main Results:
- CD271+ cells from G-CSF-mobilized PB did not yield MSC outgrowth.
- CD271+ cells from BM successfully formed MSC colonies.
- G-CSF-mobilized PB's CD271+ cells had higher hematopoietic marker expression (CD45, CD34, CD133).
Conclusions:
- CD271 enrichment is ineffective for isolating MSCs from G-CSF-mobilized PB.
- The majority of CD271+ cells in mobilized PB are hematopoietic, with a low MSC frequency.
- Current isolation methods likely result in loss of the rare MSCs during processing.

