In membrane bioreactors the cells are isolated from the bulk medium through a semipermeable membrane. This concept, which is analogous to how the circulatory system supplies solid tissues with nutrients, allows the maintenance of cells at much higher densities than is possible in traditional cultures. The membrane-based microbioreactor described herein is easy to operate, requiring only a pipette to load and harvest cells. A 10 μL culture volume was isolated from 1 mL of bulk medium through a semipermeable membrane having a molecular weight cutoff of 10 kDa. Here we describe the benefits regarding the retention of both cells and their secretions within this small culture volume using the following two model systems: hematopoietic stem cell expansion and mesenchymal stem cell-derived cartilage matrix accumulation.
Get full access to this article
View all access options for this article.
References
1.
KnazekR.A., KohlerP.O., GullinoP.M., DedrickR.L.Cell-culture on artificial capillaries—approach to tissue growth in vitro. Science, 178:65. 1972.
2.
AltshulerG.L., DziewulskiD.M., SowekJ.A., BelfortG.Continuous hybridoma growth and monoclonal-antibody production in hollow fiber reactors separators. Biotechnol Bioeng, 28:646. 1986.
3.
EvansT.L., MillerR.A.Large-scale production of murine monoclonal-antibodies using hollow fiber bioreactors. Biotechniques, 6:762. 1988.
4.
MaloneC.C., SchiltzP.M., MackintoshA.D., BeutelL.D., HeinemannF.S., DillmanR.O.Characterization of human tumor-infiltrating lymphocytes expanded in hollow-fiber bioreactors for immunotherapy of cancer. Cancer Biother Radiopharm, 16:381. 2001.
5.
HillmanG.G., WolfM.L., MontecilloE., YounesE., AliE., PontesJ.E., HaasG.P.Expansion of activated lymphocytes obtained from renal-cell carcinoma in an automated hollow-fiber bioreactor. Cell Transplant, 3:263. 1994.
6.
ShatfordR.A., NybergS.L., MeierS.J., WhiteJ.G., PayneW.D., HuW.S., CerraF.B.Hepatocyte function in a hollow fiber bioreactor—a potential bioartificial liver. J Surg Res, 53:549. 1992.
TillesA.W., BaskaranH., RoyP., YarmushM.L., TonerM.Effects of oxygenation and flow on the viability and function of rat hepatocytes cocultured in a microchannel flat-plate bioreactor. Biotechnol Bioeng, 73:379. 2001.
9.
ChanC., BerthiaumeF., NathB.D., TillesA.W., TonerM., YarmushM.L.Hepatic tissue engineering for adjunct and temporary liver support: critical technologies. Liver Transplant, 10:1331. 2004.
KimL., TohY.C., VoldmanJ., YuH.A practical guide to microfluidic perfusion culture of adherent mammalian cells. Lab Chip, 7:681. 2007.
16.
De LeonA., MayaniH., RamirezO.T.Design, characterization and application of a minibioreactor for the culture of human hematopoietic cells under controlled conditions. Cytotechnology, 28:127. 1998.
17.
GluckmanE., RochaV., BoyerChammardA., LocatelliF., ArceseW., PasquiniR., OrtegaJ., SouilletG., FerreiraE., LaporteJ.P., FernandezM., ChastangC.Outcome of cord-blood transplantation from related and unrelated donors. N Engl J Med, 337:373. 1997.
18.
WagnerJ.E., RosenthalJ., SweetmanR., ShuX.O., DaviesS.M., RamsayN.K.C., McGlaveP.B., SenderL., CairoM.S.Successful transplantation of HLA-matched and HLA-mismatched umbilical cord blood from unrelated donors: analysis of engraftment and acute graft-versus-host disease. Blood, 88:795. 1996.
19.
RobinsonS., NiuT., de LimaM., NgJ., YangH., McMannisJ., KarandishS., SadeghiT., FuP., del AngelM., O'ConnorS., ChamplinR., ShpallE.Ex vivo expansion of umbilical cord blood. Cytotherapy, 7:243. 2005.
RadisicM., DeenW., LangerR., Vunjak-NovakovicG.Mathematical model of oxygen distribution in engineered cardiac tissue with parallel channel array perfused with culture medium containing oxygen carriers. Am J Physiol Heart Circ Physiol, 288:H1278. 2005.
22.
BouhassiraE.E.Toward the manufacture of red blood cells?Blood, 112:4362. 2008.
23.
O'DriscollS.W.The healing and regeneration of articular cartilage. J Bone Joint Surg Am, 80A:1795. 1998.
24.
ChenF.H., RouscheK.T., TuanR.S.Technology insight: adult stem cells in cartilage regeneration and tissue engineering. Nat Clin Pract Rheumatol, 2:373. 2006.
BentleyG., BiantL., CarringtonR.A prospective, randomised comparison of autologous chondrocyte implantation versus mosaicplasty for osteochondral defects in the knee. J Bone Joint Surg Br, 85:223. 2003.
28.
BrittbergM., LindahlA., NilssonA., OhlssonC., IsakssonO., PetersonL.Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N Engl J Med, 331:889. 1994.
29.
van der KraanP.M., BumaP., van KuppeveltT., van den BergW.B.Interaction of chondrocytes, extracellular matrix and growth factors: relevance for articular cartilage tissue engineering. Osteoarthritis Cartilage, 10:631. 2002.
30.
DuffyD.C., McDonaldJ.C., SchuellerO.J.A., WhitesidesG.M.Rapid prototyping of microfluidic systems in poly(dimethylsiloxane)Anal Chem, 70:4974. 1998.
31.
MuschlerG.E., NakamotoC., GriffithL.G.Engineering principles of clinical cell-based tissue engineering. J Bone Joint Surg Am, 86A:1541. 2004.
32.
CollinsP.C., NielsenL.K., WongC.K., PapoutsakisE.T., MillerW.M.Real-time method for determining the colony-forming cell content of human hematopoietic cell cultures. Biotechnol Bioeng, 55:693. 1997.
33.
HinrichsW.L.J., tenHoopenH.W.M., EngbersG.H.M., FeijenJ.In vitro evaluation of heparinized Cuprophan hemodialysis membranes. J Biomed Mater Res, 35:443. 1997.
34.
BaltzJ.M., BiggersJ.D.Oxygen-transport to embryos in microdrop cultures. Mol Reprod Dev, 28:351. 1991.
35.
KirkpatrickJ.P., BrizelD.M., DewhirstM.W.A mathematical model of tumor oxygen and glucose mass transport and metabolism with complex reaction kinetics. Radiat Res, 159:336. 2003.
36.
ChowD.C., WenningL.A., MillerW.M., PapoutsakisE.T.Modeling pO(2) distributions in the bone marrow hematopoietic compartment. I. Krogh's model. Biophys J, 81:675. 2001.
37.
CwirlaS.E., BalasubramanianP., DuffinD.J., WagstromC.R., GatesC.M., SingerS.C., DavisA.M., TansikR.L., MattheakisL.C., BoytosC.M., SchatzP.J., BaccanariD.P., WrightonN.C., BarrettR.W., DowerW.J.Peptide agonist of the thrombopoietin receptor as potent as the natural cytokine. Science, 276:1696. 1997.
38.
BrookeG., TongH., LevesqueJ.P., AtkinsonK.Molecular trafficking mechanisms of multipotent mesenchymal stem cells derived from human bone marrow and placenta. Stem Cells Dev, 17:929. 2008.
39.
PrinceH.M., SimmonsP.J., WhittyG., WallD.P., BarberL., TonerG.C., SeymourJ.F., RichardsonG., MrongoviusR., HaylockD.N.Improved haematopoietic recovery following transplantation with ex vivo-expanded mobilized blood cells. Br J Haematol, 126:536. 2004.
40.
CaseJ., RiceA., WoodJ., GaudryL., VowelsM., NordonR.E.Characterization of cytokine interactions by flow cytometry and factorial analysis. Cytometry, 43:69. 2001.
41.
WognumA.W., EavesA.C., ThomasT.E.Identification and isolation of hematopoietic stem cells. Arch Med Res, 34:461. 2003.
42.
KisidayJ.D., JinM.S., DiMiccoM.A., KurzB., GrodzinskyA.J.Effects of dynamic compressive loading on chondrocyte biosynthesis in self-assembling peptide scaffolds. J Biomech, 37:595. 2004.
43.
SainiS., WickT.M.Effect of low oxygen tension on tissue-engineered cartilage construct development in the concentric cylinder bioreactor. Tissue Eng, 10:825. 2004.
