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Isolation and characterization of endothelial progenitor cells from Rhesus monkeys.

Sun W, Zheng L, Han P, Kang YJ - Regen Med Res (2014)

Bottom Line: The results showed that nonselective mononuclear EPCs were a better choice for high yield of the target cells.The cells grew in M 200 better than in EGM-2, and supplementation with fetal bovine serum promoted cell proliferation; but serum level at 7.5% was better than at 10%.This procedure would help using these valuable cells for regenerative medicine research.

View Article: PubMed Central - PubMed

Affiliation: Regenerative Medicine Research Center, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041 China.

ABSTRACT

Background: Endothelial progenitor cells (EPCs) are increasingly becoming a major focus of regenerative medicine research and practice. The present study was undertaken to establish an appropriate procedure for isolation and characterization of EPCs from Rhesus monkeys for regenerative medicine research.

Result: Selective CD34+ and nonselective mononuclear EPCs were isolated from bone marrow and cultured under varying conditions. The results showed that nonselective mononuclear EPCs were a better choice for high yield of the target cells. The cells grew in M 200 better than in EGM-2, and supplementation with fetal bovine serum promoted cell proliferation; but serum level at 7.5% was better than at 10%. In addition, surface coating of the culture dishes with human fibronectin significantly improved the proliferation and ontogeny of the isolated EPCs. Immunocytochemistry including detection of markers CD34, CD133 and CD31 and double-staining for Ac-LDL and lectin verified the purity of the cultured mononuclear EPCs.

Conclusion: By a thorough analysis, we established a practical procedure for isolation and propagation of EPCs from Rhesus monkeys. This procedure would help using these valuable cells for regenerative medicine research.

No MeSH data available.


Related in: MedlinePlus

Effects of culture dish surface coating on EPCs in cultures. Unselected mononuclear cells were seeded in EGM-2 medium without serum supplementation on FN coated culture dishes. Morphology of EPCs was respectively observed on day 5 and day 8 after culturing. In the control group, cells were seeded on culture dishes without coating. Cells seeded on FN coated dishes had better adherence than cells seeded on dishes without coating 5 days after culturing, which remained observable until 8 days after culturing. Bar =100 μm.
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Fig3: Effects of culture dish surface coating on EPCs in cultures. Unselected mononuclear cells were seeded in EGM-2 medium without serum supplementation on FN coated culture dishes. Morphology of EPCs was respectively observed on day 5 and day 8 after culturing. In the control group, cells were seeded on culture dishes without coating. Cells seeded on FN coated dishes had better adherence than cells seeded on dishes without coating 5 days after culturing, which remained observable until 8 days after culturing. Bar =100 μm.

Mentions: The unselected mononuclear EPCs were plated on culture dishes without coating or coated with 25 μg/ml FN, as shown in Figure 3. On the 5th day, cells seeded on FN-coated dishes adhered much better than cells cultured in dishes without coating, which remained observable until on the 8th day. Comparing the effects between FBS supplementation and FN coating, we found that the effect of FN coating was similar with FBS supplementation, but FBS jeopardizes the multipotential of stem cells [25].Figure 3


Isolation and characterization of endothelial progenitor cells from Rhesus monkeys.

Sun W, Zheng L, Han P, Kang YJ - Regen Med Res (2014)

Effects of culture dish surface coating on EPCs in cultures. Unselected mononuclear cells were seeded in EGM-2 medium without serum supplementation on FN coated culture dishes. Morphology of EPCs was respectively observed on day 5 and day 8 after culturing. In the control group, cells were seeded on culture dishes without coating. Cells seeded on FN coated dishes had better adherence than cells seeded on dishes without coating 5 days after culturing, which remained observable until 8 days after culturing. Bar =100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4389970&req=5

Fig3: Effects of culture dish surface coating on EPCs in cultures. Unselected mononuclear cells were seeded in EGM-2 medium without serum supplementation on FN coated culture dishes. Morphology of EPCs was respectively observed on day 5 and day 8 after culturing. In the control group, cells were seeded on culture dishes without coating. Cells seeded on FN coated dishes had better adherence than cells seeded on dishes without coating 5 days after culturing, which remained observable until 8 days after culturing. Bar =100 μm.
Mentions: The unselected mononuclear EPCs were plated on culture dishes without coating or coated with 25 μg/ml FN, as shown in Figure 3. On the 5th day, cells seeded on FN-coated dishes adhered much better than cells cultured in dishes without coating, which remained observable until on the 8th day. Comparing the effects between FBS supplementation and FN coating, we found that the effect of FN coating was similar with FBS supplementation, but FBS jeopardizes the multipotential of stem cells [25].Figure 3

Bottom Line: The results showed that nonselective mononuclear EPCs were a better choice for high yield of the target cells.The cells grew in M 200 better than in EGM-2, and supplementation with fetal bovine serum promoted cell proliferation; but serum level at 7.5% was better than at 10%.This procedure would help using these valuable cells for regenerative medicine research.

View Article: PubMed Central - PubMed

Affiliation: Regenerative Medicine Research Center, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041 China.

ABSTRACT

Background: Endothelial progenitor cells (EPCs) are increasingly becoming a major focus of regenerative medicine research and practice. The present study was undertaken to establish an appropriate procedure for isolation and characterization of EPCs from Rhesus monkeys for regenerative medicine research.

Result: Selective CD34+ and nonselective mononuclear EPCs were isolated from bone marrow and cultured under varying conditions. The results showed that nonselective mononuclear EPCs were a better choice for high yield of the target cells. The cells grew in M 200 better than in EGM-2, and supplementation with fetal bovine serum promoted cell proliferation; but serum level at 7.5% was better than at 10%. In addition, surface coating of the culture dishes with human fibronectin significantly improved the proliferation and ontogeny of the isolated EPCs. Immunocytochemistry including detection of markers CD34, CD133 and CD31 and double-staining for Ac-LDL and lectin verified the purity of the cultured mononuclear EPCs.

Conclusion: By a thorough analysis, we established a practical procedure for isolation and propagation of EPCs from Rhesus monkeys. This procedure would help using these valuable cells for regenerative medicine research.

No MeSH data available.


Related in: MedlinePlus