Open AccessEx Vivo Large‐Scale Generation of Human Platelets from Cord Blood CD34 + Cells
Author(s) -
Matsunaga Takuya,
Tanaka Ikuta,
Kobune Masayoshi,
Kawano Yutaka,
Tanaka Maki,
Kuribayashi Kageaki,
Iyama Satoshi,
Sato Tsutomu,
Sato Yasushi,
Takimoto Rishu,
Takayama Tetsuji,
Kato Junji,
Ninomiya Takafumi,
Hamada Hirofumi,
Niitsu Yoshiro
Publication year - 2006
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2006-0309
Subject(s) - biology , ex vivo , platelet , cord blood , cd34 , microbiology and biotechnology , stem cell , immunology , in vivo , genetics
In the present investigation, we generated platelets (PLTs) from cord blood (CB) CD34 + cells using a three‐phase culture system. We first cultured 500 CB CD34 + cells on telomerase gene‐transduced human stromal cells ( hTERT stroma) in serum‐free medium supplemented with stem cell factor (SCF), Flt‐3/Flk‐2 ligand (FL), and thrombopoietin (TPO) for 14 days. We then transferred the cells to hTERT stroma and cultured for another 14 days with fresh medium containing interleukin‐11 (IL‐11) in addition to the original cytokine cocktail. Subsequently, we cultured the cells in a liquid culture medium containing SCF, FL, TPO, and IL‐11 for another 5 days to recover PLT fractions from the supernatant, which were then gel‐filtered to purify the PLTs. The calculated yield of PLTs from 1.0 unit of CB (5 × 10 6 CD34 + cells) was 1.26 × 10 11 −1.68 × 10 11 PLTs. These numbers of PLTs are equivalent to 2.5–3.4 units of random donor‐derived PLTs or 2/5–6/10 of single‐apheresis PLTs. The CB‐derived PLTs exhibited features quite similar to those from peripheral blood in morphology, as revealed by electron micrographs, and in function, as revealed by fibrinogen/ADP aggregation, with the appearance of P‐selectin and activated glycoprotein IIb‐IIIa antigens. Thus, this culture system may be applicable for large‐scale generation of PLTs for future clinical use.