PMID- 33491764
OWN - NLM
STAT- MEDLINE
DCOM- 20220328
LR  - 20220328
IS  - 1097-0290 (Electronic)
IS  - 0006-3592 (Linking)
VI  - 118
IP  - 5
DP  - 2021 May
TI  - Red cell manufacturing using parallel stirred-tank bioreactors at the final 
      stages of differentiation enhances reticulocyte maturation.
PG  - 1763-1778
LID - 10.1002/bit.27691 [doi]
AB  - The aim of this study was to develop a robust, quality controlled, and 
      reproducible erythroid culture system to obtain high numbers of mature 
      erythroblasts and red blood cells (RBCs). This was achieved using a fully 
      controlled stirred-tank bioreactor by the design of experiments (DOE) methods in 
      the serum-free medium by defining the appropriate culture parameters. Human cord 
      blood CD34+ cells were first cultured in static flasks and then inoculated to 
      stirred-tank bioreactors. Cell diameter was gradually decreased and final RBC 
      yields were significantly higher when cells were inoculated at sizes smaller than 
      12 mum. The larger immature cells in the basophilic stage did not survive, while 
      smaller mature erythroid cells were successfully expanded at high agitation 
      speeds, demonstrating that appropriate seeding timing is critical. A high 
      inoculation cell density of 5 x 10(6)  cells/ml was achieved reaching 1.5 x 10(7) 
       cells/ml. By using DOE analysis fitted to precise stages of erythropoiesis, we 
      were able to acquire the optimal culture parameters for pH (7.5), temperature 
      (37 degrees C), dissolved oxygen, agitation speed (500 rpm), inoculation timing (cell 
      diameter 12-13 mum), media feeding regimen, and cell seeding density (5 x 10(6) 
       cells/ml). The final pure RBCs showed appropriate functions compared with fresh 
      donor RBCs, confirming that manufacturing mature RBCs with reproducibility is 
      possible.
CI  - (c) 2021 Wiley Periodicals LLC.
FAU - Han, So Yeon
AU  - Han SY
AD  - Department of Translational Medicine, Graduate School of Biomedical Science and 
      Engineering, Hanyang University, Seoul, Republic of Korea.
AD  - Department of Laboratory Medicine, College of Medicine, Hanyang University, 
      Seoul, Republic of Korea.
FAU - Lee, Eun Mi
AU  - Lee EM
AD  - Department of Translational Medicine, Graduate School of Biomedical Science and 
      Engineering, Hanyang University, Seoul, Republic of Korea.
AD  - Department of Laboratory Medicine, College of Medicine, Hanyang University, 
      Seoul, Republic of Korea.
FAU - Lee, Janghan
AU  - Lee J
AD  - New Drug Development Center, Osong Medical Innovation Foundation, Cheongju, 
      Republic of Korea.
AD  - College of Pharmacy, Chungbuk National University, Cheongju, Republic of Korea.
FAU - Lee, Hyosang
AU  - Lee H
AD  - New Drug Development Center, Osong Medical Innovation Foundation, Cheongju, 
      Republic of Korea.
FAU - Kwon, Amy M
AU  - Kwon AM
AUID- ORCID: 0000-0003-1632-0677
AD  - Biostatistical Consulting and Research Laboratory, Medical Research Collaborating 
      Center, Industry-University Cooperation Foundation, Hanyang University, Seoul, 
      Republic of Korea.
AD  - Biostatistics Core, Medicine-Engineering-Bio (MEB) Center, Industry-University 
      Cooperation Foundation, Hanyang University, Seoul, Republic of Korea.
FAU - Ryu, Ki Young
AU  - Ryu KY
AD  - Departmemt of Obstetrics and Gynecology, College of Medicine, Hanyang University, 
      Seoul, Republic of Korea.
FAU - Choi, Won-Seok
AU  - Choi WS
AD  - New Drug Development Center, Osong Medical Innovation Foundation, Cheongju, 
      Republic of Korea.
FAU - Baek, Eun Jung
AU  - Baek EJ
AUID- ORCID: 0000-0001-5477-6486
AD  - Department of Translational Medicine, Graduate School of Biomedical Science and 
      Engineering, Hanyang University, Seoul, Republic of Korea.
AD  - Department of Laboratory Medicine, College of Medicine, Hanyang University, 
      Seoul, Republic of Korea.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210202
PL  - United States
TA  - Biotechnol Bioeng
JT  - Biotechnology and bioengineering
JID - 7502021
SB  - IM
MH  - *Bioreactors
MH  - Cell Culture Techniques/*methods
MH  - Cell Differentiation
MH  - Cells, Cultured
MH  - Erythrocytes/*cytology
MH  - Female
MH  - Fetal Blood/cytology
MH  - Humans
MH  - Pregnancy
MH  - Reticulocytes/*cytology
OTO - NOTNLM
OT  - bioreactor
OT  - blood
OT  - design of experiments
OT  - erythrocyte
OT  - in vitro cell culture
EDAT- 2021/01/26 06:00
MHDA- 2022/03/29 06:00
CRDT- 2021/01/25 08:43
PHST- 2021/01/20 00:00 [revised]
PHST- 2020/09/01 00:00 [received]
PHST- 2021/01/22 00:00 [accepted]
PHST- 2021/01/26 06:00 [pubmed]
PHST- 2022/03/29 06:00 [medline]
PHST- 2021/01/25 08:43 [entrez]
AID - 10.1002/bit.27691 [doi]
PST - ppublish
SO  - Biotechnol Bioeng. 2021 May;118(5):1763-1778. doi: 10.1002/bit.27691. Epub 2021 
      Feb 2.