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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.In addition, surface coating of the culture dishes with human fibronectin significantly improved the proliferation and ontogeny of the isolated EPCs.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 serum concentrations on EPCs in cultures. Unselected mononuclear cells were cultured in M 200 or M 200 supplemented with 7.5%or 10%FBS,observed on day 3,day 6 and day 9. Cells in the medium supplemented with FBS grew faster than in medium without FBS supplementation, but cells in the medium supplemented with 7.5% FBS proliferated better than in the medium supplemented with 10% FBS. Bar =100 μm.
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Fig2: Effects of serum concentrations on EPCs in cultures. Unselected mononuclear cells were cultured in M 200 or M 200 supplemented with 7.5%or 10%FBS,observed on day 3,day 6 and day 9. Cells in the medium supplemented with FBS grew faster than in medium without FBS supplementation, but cells in the medium supplemented with 7.5% FBS proliferated better than in the medium supplemented with 10% FBS. Bar =100 μm.

Mentions: The unselected mononuclear EPCs were cultured in media supplemented with varying concentrations of FBS, as shown in Figure 2. On the third day, cells in the media supplemented with FBS grew faster than in media without FBS supplementation, and this phenomenon was more obvious on the 6th day and last until the 9th day. However, cells cultured in media containing 7.5% FBS proliferated better than in media supplemented with 10% FBS.Figure 2


Isolation and characterization of endothelial progenitor cells from Rhesus monkeys.

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

Effects of serum concentrations on EPCs in cultures. Unselected mononuclear cells were cultured in M 200 or M 200 supplemented with 7.5%or 10%FBS,observed on day 3,day 6 and day 9. Cells in the medium supplemented with FBS grew faster than in medium without FBS supplementation, but cells in the medium supplemented with 7.5% FBS proliferated better than in the medium supplemented with 10% FBS. Bar =100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig2: Effects of serum concentrations on EPCs in cultures. Unselected mononuclear cells were cultured in M 200 or M 200 supplemented with 7.5%or 10%FBS,observed on day 3,day 6 and day 9. Cells in the medium supplemented with FBS grew faster than in medium without FBS supplementation, but cells in the medium supplemented with 7.5% FBS proliferated better than in the medium supplemented with 10% FBS. Bar =100 μm.
Mentions: The unselected mononuclear EPCs were cultured in media supplemented with varying concentrations of FBS, as shown in Figure 2. On the third day, cells in the media supplemented with FBS grew faster than in media without FBS supplementation, and this phenomenon was more obvious on the 6th day and last until the 9th day. However, cells cultured in media containing 7.5% FBS proliferated better than in media supplemented with 10% FBS.Figure 2

Bottom Line: The results showed that nonselective mononuclear EPCs were a better choice for high yield of the target cells.In addition, surface coating of the culture dishes with human fibronectin significantly improved the proliferation and ontogeny of the isolated EPCs.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