Ex vivo expansion and clonality of CD34+ selected cells from bone marrow and cord blood in a serum-free media.
Although umbilical cord blood is increasingly being used in allogeneic marrow transplantation, delayed platelet engraftment is often a concern for cord blood transplant recipients. We evaluated the potential of ex vivo expansion and clonality in CD34+ cells separated from a bone marrow source, and cord blood, in a serum-free Media. The CD34+ cells, selected from bone marrow (BM) and umbilical cord blood (CB), were expanded with hematopoietic growth factors. They were then cultured for burst-forming units of erythrocytes (BFU-E), colony-forming units of granulocytes and monocytes (CFU-GM) and colony-forming units of megakaryocytes (CFU-Mk) at days 0, 4, 7, and 14 under the combination of growth factors, with cell counts. The cytokines included the recombinant human megakaryocyte growth and development (100 ng/ml), interleukin-3 (10 ng/ml), stem cell factor (100 ng/ml), flt-3 ligand (50 ng/ml) and interleukin-11 (200 ng/ml). The CB-selected CD34+ cells showed significantly higher total cell expansion than those from the BM at day 7 (3.0 fold increase than BM), day 14 (2.4 fold), and day 17 (2.6 fold). The colony count of the BFU-E/CFU-E per CD34+ cell at day 0 was 0.14 +/- 0.023 in the CB, which was significantly higher than 0.071 +/- 0.015 in the BM. The CB-selected CD34+ cells produced more BFU-E colonies than the BM on culture days 4, 7, and 14. The BFU-E colonies from the CB cells increased markedly on culture days 4 and 7, with a 4-fold increase at day 14. The colony count of the CFU-Mk per CD34+ cell at day 0 was 0.047 +/- 0.011 in the CB-selected CD34+ cells cultures, which was higher than the 0.026 +/- 0.014 in the BM. The CB-selected CD34+ cells produced more CFU-Mk colonies than the BM on culture days 4, 7 and 14. In conclusion, the ex vivo expansion of the CB cells may be very promising in producing total cellular expansion, CFU-Mk and BFU-E compared with BM, especially at day 7. The ex vivo expansion of the CB may have rationale in making an ex vivo culture for 7 to 14 d.
Kim
Soon-Ki
SK
Department of Pediatrics, Inha University of Hospital, Inha Medical College, Inchon 402-751, Korea. pedkim@inha.ac.kr
Koh
Seung-Kwon
SK
Song
Sun U
SU
Shin
Seok-Hwan
SH
Choi
Gwang-Seong
GS
Kim
Woo-Chul
WC
Lee
Moon-Hee
MH
Seoh
Ju Young
JY
Park
San Kyu
SK
Fraser
John K
JK
D018952:Antigens, CD34 / Q000378:metabolism*; D001772:Blood Cell Count; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002469:Cell Separation; D002999:Clone Cells / Q000378:metabolism; D003114:Colony-Forming Units Assay; D016895:Culture Media, Serum-Free; D016207:Cytokines / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism; D006133:Growth Substances / Q000378:metabolism; D006801:Humans
D018952:Antigens, CD34; D016895:Culture Media, Serum-Free; D016207:Cytokines; D006133:Growth Substances