title: Microliter-bioreactor array with buoyancy-driven stirring for human hematopoietic stem cell culture.
abstract: This work presents the development of an array of bioreactors where finely controlled stirring is provided at the microliter scale (100-300 mul). The microliter-bioreactor array is useful for performing protocol optimization in up to 96 parallel experiments of hematopoietic stem cell (HSC) cultures. Exploring a wide range of experimental conditions at the microliter scale minimizes cost and labor. Once the cell culture protocol is optimized, it can be applied to large-scale bioreactors for stem cell production at the clinical level. The controlled stirring inside the wells of a standard 96-well plate is provided by buoyancy-driven thermoconvection. The temperature and velocity fields within the culture volume are determined with numerical simulations. The numerical results are verified with experimental velocity measurements using microparticle image velocimetry (muPIV) and are used to define feasible experimental conditions for stem cell cultures. To test the bioreactor array's functionality, human umbilical cord blood-derived CD34(+) cells were cultured for 7 days at five different stirring conditions (0.24-0.58 mums) in six repeated experiments. Cells were characterized in terms of proliferation, and flow cytometry measurements of viability and CD34 expression. The microliter-bioreactor array demonstrates its ability to support HSC cultures under stirred conditions without adversely affecting the cell behavior. Because of the highly controlled operative conditions, it can be used to explore culture conditions where the mass transport of endogenous and exogenous growth factors is selectively enhanced, and cell suspension provided. While the bioreactor array was developed for culturing HSCs, its application can be extended to other cell types.
authors.forename: Camilla
authors.initials: C
authors.lastname: Luni
authors.forename: Hope C
authors.initials: HC
authors.lastname: Feldman
authors.forename: Michela
authors.initials: M
authors.lastname: Pozzobon
authors.forename: Paolo
authors.initials: P
authors.lastname: De Coppi
authors.forename: Carl D
authors.initials: CD
authors.lastname: Meinhart
authors.forename: Nicola
authors.initials: N
authors.lastname: Elvassore
references.citation: Lab Hematol. 2006;12(2):86-93
references.pmid: 16751136
references.citation: Cell Stem Cell. 2008 Oct 9;3(4):369-81
references.pmid: 18940729
references.citation: Biotechnol Prog. 1998 May-Jun;14(3):466-72
references.pmid: 9622528
references.citation: J Cell Physiol. 1977 Jun;91(3):335-44
references.pmid: 301143
references.citation: Bone Marrow Transplant. 2007 Jan;39(1):11-23
references.pmid: 17164824
references.citation: Methods Enzymol. 2006;419:439-73
references.pmid: 17141066
references.citation: J Clin Apher. 2001;16(4):186-91
references.pmid: 11835415
references.citation: Cytotechnology. 2008 Mar;56(3):151-60
references.pmid: 19002853
references.citation: Bone Marrow Transplant. 2006 Jul;38(2):83-93
references.pmid: 16751788
references.citation: Trends Biotechnol. 2003 May;21(5):233-40
references.pmid: 12727385
references.citation: Biotechnology (N Y). 1994 Sep;12(9):909-14
references.pmid: 7521647
references.citation: Blood. 1992 Jun 1;79(11):2821-6
references.pmid: 1375113
references.citation: Nature. 2006 Jun 29;441(7097):1075-9
references.pmid: 16810242
references.citation: Annu Rev Biomed Eng. 1999;1:129-52
references.pmid: 11701485
references.citation: N Engl J Med. 2006 Sep 7;355(10):1070; author reply 1070-1
references.pmid: 16957160
references.citation: Am J Pathol. 2006 Aug;169(2):338-46
references.pmid: 16877336
references.citation: Lab Chip. 2007 Jun;7(6):710-9
references.pmid: 17538712
references.citation: Springer Semin Immunopathol. 2004 Nov;26(1-2):143-54
references.pmid: 15549306
references.citation: Cytotechnology. 1998 Nov;28(1-3):127-38
references.pmid: 19003414
references.citation: Exp Hematol. 2008 Feb;36(2):235-43
references.pmid: 18206730
references.citation: Cryobiology. 2003 Feb;46(1):61-75
references.pmid: 12623029
references.citation: J Biomed Mater Res. 1997 Sep 5;36(3):347-59
references.pmid: 9260106
references.citation: Biotechnol Prog. 1993 Mar-Apr;9(2):131-7
references.pmid: 7763589