Method and Apparatus for a Miniature Bioreactor System for Long-Term Cell Culture

48 Pages·2015·5.14 MB·English

https://ntrs.nasa.gov/search.jsp?R=20150009343 2019-04-12T03:39:29+00:00Z
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(12) United States Patent (1o) Patent No.: (cid:9) US 9,023,642 B2
Kleis et al. (45) Date of Patent: (cid:9) May 5, 2015
(54) METHOD AND APPARATUS FOR A (56) References Cited
MINIATURE BIOREACTOR SYSTEM FOR
LONG-TERM CELL CULTURE U.S. PATENT DOCUMENTS
(75) Inventors: Stanley J. Kleis, Houston, TX (US); 5,153,131 A 10/1992 Wolf et al.
Sandra K. Geffert, Kingwood, TX 5,496,722 A 3/1996 Goodwin et al.
5,763,279 A 6/1998 Schwarz et al.
(US); Steve R. Gonda, Houston, TX
6,120,735 A * 9/2000 Zborowski et al . (cid:9) ............. (cid:9) 422/73
(US) 6,632,619 131 * 10/2003 Harrison et al . (cid:9) ............... (cid:9) 435/7.2
6,867,040 132 3/2005 Helmstetter et al.
(73) Assignee: The University of Houston System,
2006/0172921 Al* 8/2006 Freyberg et al . (cid:9) .................. (cid:9) 514/2
Houston, TX (US)
OTHER PUBLICATIONS
(*) Notice: (cid:9) Subject to any disclaimer, the term of this
patent is extended or adjusted under 35 Goodwin, T.J., Jessup JM, Wolf, DA, Spaulding GF, 1993. Reduced
U.S.C. 154(b) by 1607 days. Shear Stress: a major component in the ability ofmammalian tissues
to form three-dimensional assemblies in simulated microgravity. J
(21) Appl. No.: 11/774,550
Cell Biochem 51:Abstract Only
(22) Filed: (cid:9) Jul. 6, 2007 Levesque, M.J. and Nerem, R.M. 1985, The elongation and orien-
tation of cultured endothelial cells in response to shear stress. Journal
(65) (cid:9) Prior Publication Data of Biomechanical Engineering. vol. 107, pp.Abstract Only.
US 2008/0032380 Al (cid:9) Feb. 7, 2008
* cited by examiner
Related U.S. Application Data
Primary Examiner William H Beisner
(60) Provisional application No. 60/819,325, filed on Jul. 7,
(74) Attorney, Agent, or Firm Robert W Strozier
2006.
(51) Int. Cl. (57) ABSTRACT
C12M 3104 (cid:9) (2006.01) A bioreactor and method that permits continuous and simul-
C12N 5100 (cid:9) (2006.01) taneous short, moderate, or long term cell culturing of one or
C12M 1/00 (cid:9) (2006.01) more cell types or tissue in a laminar flow configuration is
C12M 1112 (cid:9) (2006.01) disclosed, where the bioreactor supports at least two laminar
C12M 1104 (cid:9) (2006.01) flow zones, which are isolated by laminar flow without the
(52) U.S. Cl. need for physical barriers between the zones. The bioreactors
CPC ............... C12M 29/10 (2013.01); C12M 23/04 of this invention are ideally suited for studying short, moder-
(2013.01); C12M 23124 (2013.01); C12M 29120 ate and long term studies of cell cultures and the response of
(2013.01) cell cultures to one or more stressors such as pharmaceuticals,
(58) Field of Classification Search hypoxia, pathogens, or any other stressor. The bioreactors of
CPC ...... C12M 23/04; C12M 23/24; C12M 29/10; this invention are also ideally suited for short, moderate or
C12M 29/20; BOIL 3/502776; BOIL long term cell culturing with periodic cell harvesting and/or
2200/0636; BOIL 2300/0877 medium processing for secreted cellular components.
USPC .................... 435/297.2, 299.1, 378, 395, 402
See application file for complete search history. 14 Claims, 31 Drawing Sheets
U.S. Patent (cid:9) May 5, 2015 (cid:9) Sheet I of 31 (cid:9) US 9,023,642 B2
I I
12-La 12 LL22 06 cg
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U.S. Patent (cid:9) US 9,023,642 B2
May 5, 2015 (cid:9) Sheet 2 of 31 (cid:9)
12fie (cid:9) 1 (cid:9) 126e'
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U.S. Patent (cid:9)
May 5, 2015 (cid:9) Sheet 3 of 31 (cid:9) US 9,023,642 B2
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U.S. Patent (cid:9) May 5, 2015 (cid:9) Sheet 4 of 31 (cid:9) US 9,023,642 B2
224
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U.S. Patent (cid:9) May 5, 2015 (cid:9) Sheet 5 of 31 (cid:9) US 9,023,642 B2
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U.S. Patent
May 5, 2015 (cid:9) Sheet 6 of 31 (cid:9) US 9,023,642 B2
300
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FIG.. 3
206
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FIG. 3B
U.S. Patent (cid:9) May 5, 2015 (cid:9) Sheet 7 of 31 (cid:9) US 9,023,642 B2
400
FIG, 4A
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300
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FIG, 4B
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U.S. Patent (cid:9) May 5, 2015 (cid:9) Sheet 8 of 31 (cid:9) US 9,023,642 B2
510 (cid:9)
508
5 (cid:9)
502
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U.S. Patent (cid:9) May 5, 2015 (cid:9) Sheet 9 of 31 (cid:9) US 9,023,642 B2
610 (cid:9)
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