scholarly journals Isolation of Muscle-Derived Stem/Progenitor Cells Based on Adhesion Characteristics to Collagen-Coated Surfaces

2013 ◽  
pp. 53-65 ◽  
Author(s):  
Mitra Lavasani ◽  
Aiping Lu ◽  
Seth D. Thompson ◽  
Paul D. Robbins ◽  
Johnny Huard ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Coated Surfaces

Enrichment of CD34+ Hematopoietic Stem and Progenitor Cells from Human Bone Marrow Using a P-Selectin-Coated Microtube.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1219-1219
Author(s):  
Srinivas D. Narasipura ◽  
Jane L. Liesveld ◽  
Joel C. Wojciechowski ◽  
Nichola Charles ◽  
Karen Rosell ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Adhesion Molecule ◽  
Mononuclear Cells ◽  
Single Cells ◽  
Bone Marrow Mononuclear Cells ◽  
Cell Capture ◽  
Hematopoietic Stem ◽  
Stem And Progenitor Cells ◽  
Coated Surfaces

Abstract Enrichment and purification of hematopoietic stem and progenitor cells (HSPCs) is important in transplantation therapies for hematological disorders and for basic stem cell research. Primitive CD34+ HSPCs have demonstrated stronger rolling adhesion than mature CD34- mononuclear cells on selectins (Blood2000; 95:478–486). We have exploited this differential rolling behavior to capture and purify HSPCs from bone marrow, by perfusing mononuclear cells through selectin-coated microtubes. Bone marrow mononuclear cells were perfused through the cell capture microtubes coated with adhesion molecules. These utilized a parallel plate flow chamber (Glycotech), and the P-selectin was adsorbed with laboratory tubing of appropriate lengths attached to the inlet, outlet, and vacuum ports of the gasket chamber. After perfusion, the device lumen was washed and captured cells were visualized and estimated by video microscopy. “Rolling” cells were defined as cells translating at less than 50% of the calculated hydrodynamic free stress velocity. Velocities of single cells were determined using a MATLAB program designed to measure the change in position of the cell centroid in a given time period. Adherent cells were eluted by high shear, calcium free buffer and air embolism. Immunofluorescence staining followed by flow cytometry was used to analyze CD34+ HSPCs. CD34+ HSPC purity of cells captured in adhesion molecule-coated devices was significantly higher than the fraction of CD34+ cells found in bone marrow- mononuclear cells (2.5 ± 0.8%). P-selectin coated surfaces yielded 16–20% CD34+ cell purity, while antibody coated surfaces yielded 12–18%. Although the CD34+ cell purities were comparable between selectin and antibody surfaces, the total number of CD34+ HSPCs captured was significantly higher in P-selectin devices (∼5.7–7.1 × 104) when compared to the antibody device (∼1.74–2.61 × 104). Furthermore, analysis for cells positive for CD133, a surface marker for more primitive HSPCs, depicted approximately 10–14 fold enrichment in P-selectin samples over control bone marrow mononuclear cells. The captured cells were viable and exhibited in vitro colony forming capabilities. Thus, P-selectin can be used in a compact flow device to capture and enrich HSPCs. This study supports the hypothesis that flow-based adhesion molecule-mediated capture may be a viable physiologic approach to the capture and purification of HSPCs.

scholarly journals P-Selectin–Coated Microtube for Enrichment of CD34+ Hematopoietic Stem and Progenitor Cells from Human Bone Marrow

2008 ◽  
Vol 54 (1) ◽  
pp. 77-85 ◽  
Author(s):  
Srinivas D Narasipura ◽  
Joel C Wojciechowski ◽  
Nichola Charles ◽  
Jane L Liesveld ◽  
Michael R King
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Adhesion Molecule ◽  
Mononuclear Cells ◽  
Air Embolism ◽  
Cell Capture ◽  
Hematopoietic Stem ◽  
Cd34 Cells ◽  
Stem And Progenitor Cells ◽  
Coated Surfaces

Abstract Background: Enrichment and purification of hematopoietic stem and progenitor cells (HSPCs) is important in transplantation therapies for hematologic disorders and in basic stem cell research. Primitive CD34+ HSPCs have demonstrated stronger rolling adhesion on selectins than mature CD34− mononuclear cells (MNCs). We have exploited this differential rolling behavior to capture and purify HSPCs from bone marrow by perfusing MNCs through selectin-coated microtubes. Methods: Bone marrow MNCs were perfused through the cell-capture microtubes coated with adhesion molecules. We washed the device lumen and visualized and estimated captured cells by video microscopy. Adherent cells were eluted by high shear, calcium-free buffer, and air embolism. We used immunofluorescence staining followed by flow cytometry to analyze CD34+ HSPCs. Results: CD34+ HSPC purity of cells captured in adhesion molecule–coated devices was significantly higher than the fraction of CD34+ cells found in bone marrow MNCs [mean (SE) 2.5% (0.8%)]. P-selectin–coated surfaces yielded 16% to 20% CD34+ cell purity, whereas antibody-coated surfaces yielded 12% to 18%. Although CD34+ cell purity was comparable between selectin and antibody surfaces, the total number of CD34+ HSPCs captured was significantly higher in P-selectin devices (approximately 5.7 × 104 to 7.1 × 104) than antibody devices (approximately 1.74 × 104 to 2.61 × 104). Conclusions: P-selectin can be used in a compact flow device to capture HSPCs. Selectin-mediated capture of CD34+ HSPCs resulted in enrichment approximately 8-fold higher than the CD34+ cell population from bone marrow MNCs. This study supports the hypothesis that flow-based, adhesion molecule–mediated capture may be a viable alternative approach to the capture and purification of HSPCs.

scholarly journals Isolating and Expanding Endothelial Progenitor Cells from Peripheral Blood on Peptide‐functionalized Polystyrene Surfaces

2019 ◽  
Author(s):  
Mohamed Elkhodiry ◽  
Marieve Boulanger ◽  
Omar Bashth ◽  
Jean-François Tanguay ◽  
Gaétan Laroche ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Extracellular Matrix Proteins ◽  
Matrix Proteins ◽  
Endothelial Progenitor ◽  
Coated Surfaces

The expansion of human peripheral blood endothelial progenitor cells to obtain therapeutically relevant endothelial colony forming cells (ECFCs) has been commonly performed on xeno‐derived extracellular matrix proteins. For cellular therapy applications, xeno‐free culture conditions are desirable to improve product safety and reduce process variability. We have previously described a novel fluorophore‐tagged RGD peptide (RGD‐TAMRA) that enhanced the adhesion of mature endothelial cells in vitro. To investigate whether this peptide can replace animal‐derived extracellular matrix proteins in the isolation and expansion of ECFCs, peripheral blood mononuclear cells from 22 adult healthy adult donors were seeded on RGD‐TAMRA‐modified polystyrene culture surfaces. Endothelial colony formation was significantly enhanced on RGD‐TAMRA‐modified surfaces compared to the unmodified control. No phenotypic differences were detected between ECFCs obtained on RGD‐TAMRA compared to ECFCs obtained on rat tail collagen‐coated surfaces. Compared to collagen‐coated surfaces and unmodified surfaces, RGD‐TAMRA surfaces promoted ECFC adhesion, cell spreading, and clonal expansion. This work presents a platform that allows for a comprehensive in vitro evaluation of peptide‐based biofunctionalization as a promising avenue for ex vivo ECFC expansion.

Regulatory effect of dimethylarginine dimethylaminohydrolase on endothelial progenitor cells differentiation and function

2010 ◽  
Vol 34 (8) ◽  
pp. S42-S42
Author(s):  
Qiong Yuan ◽  
Chang‑Ping Hu ◽  
Si‑Yu Liu ◽  
Xu‑Meng Chen ◽  
Jun Peng ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Regulatory Effect ◽  
Dimethylarginine Dimethylaminohydrolase ◽  
Endothelial Progenitor ◽  
And Function

Wnt signaling pathway regulates the differentiation of hepatic progenitor cells

2010 ◽  
Vol 34 (8) ◽  
pp. S41-S41
Author(s):  
Yang Bi ◽  
Yun He ◽  
Tingyu Li ◽  
Tao Feng ◽  
Tongchuan He
Keyword(s):  
Wnt Signaling ◽  
Progenitor Cells ◽  
Signaling Pathway ◽  
Wnt Signaling Pathway ◽  
Hepatic Progenitor Cells

Human thymic epithelial cells maintain long-term survival of clonogenic myeloid and erythroid progenitor cells in vitro

2000 ◽  
Vol 111 (1) ◽  
pp. 363-370 ◽  
Author(s):  
Katsuto Takenaka ◽  
Mine Harada ◽  
Tomoaki Fujisaki ◽  
Koji Nagafuji ◽  
Shinichi Mizuno ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Progenitor Cells ◽  
Thymic Epithelial Cells ◽  
Erythroid Progenitor ◽  
Term Survival ◽  
Long Term Survival ◽  
Erythroid Progenitor Cells

“Blastosphere”: A new culture method for human fetal hepatic progenitor cells

2001 ◽  
Vol 120 (5) ◽  
pp. A542-A543
Author(s):  
T HIROSE ◽  
K YASUCHIKA ◽  
T FUJIKAWA ◽  
H FUJII ◽  
S OE ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Culture Method ◽  
Hepatic Progenitor Cells
Sign in / Sign up

Export Citation Format

Share Document