J Korean Med Sci.  2004 Oct;19(5):635-639. 10.3346/jkms.2004.19.5.635.

Stem Cells Expressing Homing Receptors Could be Expanded From Cryopreserved and Unselected Cord Blood

• Lee YH
• Han JY
• Seo SY
• Kim KH
• Lee YA
• Lee YS
• Lee HS
• Hur WJ
• Han H
• Kwon HC
• Kim JS
• Kim HJ

• KMID: 1733500
• DOI: http://doi.org/10.3346/jkms.2004.19.5.635

Abstract

We assessed the cytokine combinations that are best for ex vivo expansion of cord blood (CB) and the increment for cell numbers of nucleated cells, as well as stem cells expressing homing receptors, by an ex vivo expansion of cryopreserved and unselected CB. Frozen leukocyte concentrates (LC) from CB were thawed and cultured at a concentration of 1x10(5)/mL in media supplemented with a combination of SCF (20 ng/mL)+TPO (50 ng/mL)+FL (50 ng/mL)+/-IL-6 (20 ng/mL)+/-G-CSF (20 ng/mL). After culturing for 14 days, the expansion folds of cell numbers were as follows: TNC 22.3+/-7.8~26.3+/-4.9, CFU-GM 4.7+/-5.1~11.7+/-2.6, CD34+CD38- cell 214.0+/-251.9~464.1+/-566.1, CD34+CXCR4+ cell 4384.5+/-1664.7~7087.2+/-4669.3, CD34+VLA4+ cell 1444.3+/-1264.0~2074.9+/-1537.0, CD34+VLA5+ cell 86.2+/-50.9~ 113.2+/-57.1. These results revealed that the number of stem cells expressing homing receptors could be increased by an ex vivo expansion of cryopreserved and unselected CB using 3 cytokines (SCF, TPO, FL) only. Further in vivo studies regarding the engraftment after expansion of the nucleated cells, as well as the stem cells expressing homing receptors will be required.

Keyword

Receptors, Lymphocyte Homing; Ex Vivo Expansion; Cryopreservation; Fetal Blood; Cytokines; Granulocyte Colong-Stimulating Factor; Thrombopoietin; Interleukin-6: Antigen, CD34

MeSH Terms

ADP-ribosyl Cyclase/metabolism
Antigens, CD/metabolism
Antigens, CD34/metabolism
Cryopreservation
Fetal Blood/*cytology
Humans
Integrin alpha4beta1/metabolism
Integrin alpha5beta1/metabolism
Membrane Proteins
Receptors, CXCR4/metabolism
Receptors, Lymphocyte Homing/*metabolism
Research Support, Non-U.S. Gov't
Stem Cell Factor
Stem Cells/*cytology/*metabolism
Thrombopoietin

Figure

• 

  Fig. 1 The fold increases for the absolute numbers of total nucleated cells (TNC), colony forming unit-granulocyte/macrophage (CFU-GM) and CD34+CD38- cells after 14 days of culture in the presence of various cytokines. SCF, stem cell factor; TPO, thrombopoietin; FL, flt3 ligand; IL, interleukin; G-CSF, granulocyte-colony stimulating factor.

• 

  Fig. 2 The fold increases for the absolute numbers of CD34+CX CR4+, CD34+VLA4+ and CD34+VLA5+ cells after 14 days of culture in the presence of various cytokines. SCF, stem cell factor; TPO, thrombopoietin; FL, flt3 ligand; IL, interleukin; G-CSF, granulocyte-colony stimulating factor.

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