Patrick J Hanley1, Zhuyong Mei2, April G Durett2, Maria da Graca Cabreira-Hansen, Marie da Graca Cabreira-Harrison3, Mariola Klis2, Wei Li2, Yali Zhao2, Bing Yang4, Kaushik Parsha4, Osman Mir4, Farhaan Vahidy4, Debra Bloom5, R Brent Rice6, Peiman Hematti5, Sean I Savitz4, Adrian P Gee2. 1. Center for Cell and Gene Therapy, Texas Children's Hospital, The Methodist Hospital, and Baylor College of Medicine, Houston, Texas, USA; Program for Cell Enhancement and Technologies for Immunotherapy, Sheikh Zayed Institute for Pediatric Surgical Innovation, and The Center for Cancer and Immunology Research, Children's National Medical Center, Washington, DC, USA. Electronic address: phanley@childrensnational.org. 2. Center for Cell and Gene Therapy, Texas Children's Hospital, The Methodist Hospital, and Baylor College of Medicine, Houston, Texas, USA. 3. The Texas Heart Institute, St Luke's Episcopal Hospital, Houston, Texas, USA. 4. Department of Neurology, University of Texas Health Science Center, Houston, Texas, USA. 5. University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin, USA. 6. Terumo BCT, Lakewood, Colorado, USA.
Abstract
BACKGROUND: The use of bone marrow-derived mesenchymal stromal cells (MSCs) as a cellular therapy for various diseases, such as graft-versus-host disease, diabetes, ischemic cardiomyopathy and Crohn's disease, has produced promising results in early-phase clinical trials. However, for widespread application and use in later phase studies, manufacture of these cells must be cost-effective, safe and reproducible. Current methods of manufacturing in flasks or cell factories are labor-intensive, involve a large number of open procedures and require prolonged culture times. METHODS: We evaluated the Quantum Cell Expansion System for the expansion of large numbers of MSCs from unprocessed bone marrow in a functionally closed system and compared the results with a flask-based method currently in clinical trials. RESULTS: After only two passages, we were able to expand a mean of 6.6 × 10(8) MSCs from 25 mL of bone marrow reproducibly. The mean expansion time was 21 days, and cells obtained were able to differentiate into all three lineages: chondrocytes, osteoblasts and adipocytes. The Quantum was able to generate the target cell number of 2.0 × 10(8) cells in an average of 9 fewer days and in half the number of passages required during flask-based expansion. We estimated that the Quantum would involve 133 open procedures versus 54,400 in flasks when manufacturing for a clinical trial. Quantum-expanded MSCs infused into an ischemic stroke rat model were therapeutically active. CONCLUSIONS: The Quantum is a novel method of generating high numbers of MSCs in less time and at lower passages when compared with flasks. In the Quantum, the risk of contamination is substantially reduced because of the substantial decrease in open procedures.
BACKGROUND: The use of bone marrow-derived mesenchymal stromal cells (MSCs) as a cellular therapy for various diseases, such as graft-versus-host disease, diabetes, ischemic cardiomyopathy and Crohn's disease, has produced promising results in early-phase clinical trials. However, for widespread application and use in later phase studies, manufacture of these cells must be cost-effective, safe and reproducible. Current methods of manufacturing in flasks or cell factories are labor-intensive, involve a large number of open procedures and require prolonged culture times. METHODS: We evaluated the Quantum Cell Expansion System for the expansion of large numbers of MSCs from unprocessed bone marrow in a functionally closed system and compared the results with a flask-based method currently in clinical trials. RESULTS: After only two passages, we were able to expand a mean of 6.6 × 10(8) MSCs from 25 mL of bone marrow reproducibly. The mean expansion time was 21 days, and cells obtained were able to differentiate into all three lineages: chondrocytes, osteoblasts and adipocytes. The Quantum was able to generate the target cell number of 2.0 × 10(8) cells in an average of 9 fewer days and in half the number of passages required during flask-based expansion. We estimated that the Quantum would involve 133 open procedures versus 54,400 in flasks when manufacturing for a clinical trial. Quantum-expanded MSCs infused into an ischemic strokerat model were therapeutically active. CONCLUSIONS: The Quantum is a novel method of generating high numbers of MSCs in less time and at lower passages when compared with flasks. In the Quantum, the risk of contamination is substantially reduced because of the substantial decrease in open procedures.
Authors: Elena Klyushnenkova; Joseph D Mosca; Valentina Zernetkina; Manas K Majumdar; Kirstin J Beggs; Donald W Simonetti; Robert J Deans; Kevin R McIntosh Journal: J Biomed Sci Date: 2005 Impact factor: 8.410
Authors: E M Horwitz; D J Prockop; L A Fitzpatrick; W W Koo; P L Gordon; M Neel; M Sussman; P Orchard; J C Marx; R E Pyeritz; M K Brenner Journal: Nat Med Date: 1999-03 Impact factor: 53.440
Authors: Patrick J Hanley; Zhuyong Mei; Maria da Graca Cabreira-Hansen; Mariola Klis; Wei Li; Yali Zhao; April G Durett; Xingwu Zheng; Yongping Wang; Adrian P Gee; Edwin M Horwitz Journal: Cytotherapy Date: 2013-04 Impact factor: 5.414
Authors: Edwin M Horwitz; Patricia L Gordon; Winston K K Koo; Jeffrey C Marx; Michael D Neel; Rene Y McNall; Linda Muul; Ted Hofmann Journal: Proc Natl Acad Sci U S A Date: 2002-06-25 Impact factor: 11.205
Authors: Bahey Salem; Samantha Miner; Nancy F Hensel; Minoo Battiwalla; Keyvan Keyvanfar; David F Stroncek; Adrian P Gee; Patrick J Hanley; Catherine M Bollard; Sawa Ito; A John Barrett Journal: Cytotherapy Date: 2015-09-28 Impact factor: 5.414
Authors: Debra D Bloom; John M Centanni; Neehar Bhatia; Carol A Emler; Diana Drier; Glen E Leverson; David H McKenna; Adrian P Gee; Robert Lindblad; Derek J Hei; Peiman Hematti Journal: Cytotherapy Date: 2014-11-21 Impact factor: 5.414
Authors: David A Castilla-Casadiego; Ana M Reyes-Ramos; Maribella Domenech; Jorge Almodovar Journal: Ann Biomed Eng Date: 2019-11-08 Impact factor: 3.934
Authors: Ayesha Aijaz; Matthew Li; David Smith; Danika Khong; Courtney LeBlon; Owen S Fenton; Ronke M Olabisi; Steven Libutti; Jay Tischfield; Marcela V Maus; Robert Deans; Rita N Barcia; Daniel G Anderson; Jerome Ritz; Robert Preti; Biju Parekkadan Journal: Nat Biomed Eng Date: 2018-06-11 Impact factor: 25.671
Authors: John S K Yuen; Andrew J Stout; N Stephanie Kawecki; Sophia M Letcher; Sophia K Theodossiou; Julian M Cohen; Brigid M Barrick; Michael K Saad; Natalie R Rubio; Jaymie A Pietropinto; Hailey DiCindio; Sabrina W Zhang; Amy C Rowat; David L Kaplan Journal: Biomaterials Date: 2021-11-29 Impact factor: 15.304
Authors: Jonathan Sheu; Jim Beltzer; Brian Fury; Katarzyna Wilczek; Steve Tobin; Danny Falconer; Jan Nolta; Gerhard Bauer Journal: Mol Ther Methods Clin Dev Date: 2015-06-17 Impact factor: 6.698