Development of a perfusion bioreactor for culturing hematopoietic stem cells

Hematopoietic stem cells (HSC) transplantation can provide cure to a variety of malignant and non-malignant diseases. However, the success of HSC transplantation in different clinical setting largely depends on its availability. Getting acceptable quality and precise number of progenitor stem cells is difficult and in some cases impossible due to various reasons. HSC transplantation would be greatly facilitated by ex vivo expansion of a small number of cells from bone marrow, or from peripheral blood, or from a cord blood sample. At present, no suitable system is available for ex vivo expansion of stem cells. Thus, the main idea of the project is to develop a prototype bioreactor for culturing adult stem cells.

The project aims at development of a bioreactor and its operation for cultivation of hematopoietic stem cells of murine origin, mimicking bone marrow micro-environment. The specific objectives to fulfil the above mentioned aim are (i) developing a biocompatible 3-D matrix from natural and/or synthetic polymers, (ii) culture of murine HSC on 3-D matrix, pre-exposed to either stromal cells or extra cellular matrix (ECM) components, (iii) study on self-propagating potential of early progenitor cells and its differentiation into cells of myeloid lineage on support of stromal cells or its equivalent, and (iv) developing a prototype bioreactor for expansion of early progenitor stem cells and optimization of parameters for bioreactor operating.

The project has been reinitiated; the present study cover objectives (i) and (ii). Three synthetic polymers are examined in terms of substratum for growth of murine stroma cells and hematopoiesis. Further, four different ECM components are tested in adhesion assay of the progenitor stem cells. Preliminary data showed that out of three polymers, one indicated natural affinity towards bone marrow stroma-stem cells. On the other hand, all four ECM components provided support for attachment and proliferation of progenitor stem cells in stroma-free environment.

Publications

Original peer-reviewed articles

1.     ¶Anto RJ, Mukhopadhyay A, Denning K and Aggarwal BB (2002) Curcumin (diferuloylmethane) induces apoptosis through activation of caspase-8, BID cleavage and cytochrome C release: Its suppression by ectopic expression of Bcl-2 and Bcl-xL.Carcinogenesis (in press) (¶on deputation/work done elsewhere).

2.     ¶Mukhopadhyay A, Shishodia S, Fu X-Y and Aggarwal BB (2002) Lack of requirement of STAT1 for activation of NF-kB, c-jun N-terminal protein kinase, and apoptosis by TNFa. J Cellular Biochem. (in press) (¶on deputation/work done elsewhere).

3.     ¶Mukhopadhyay A, Bueso-Ramos C, Chatterjee D, Pantazis P and Aggarwal BB (2001) Curcumin downregulates cell survival mechanisms in human prostate cancer cell lines. Oncogene 20:5797-5809 (¶on deputation/work done elsewhere).

4.     ¶Mukhopadhyay A, Suttles J, Stout RD and Aggarwal BB (2001) Genetic deletion of the tumor necrosis factor receptor p60 or p80 abrogates ligand-mediated activation of nuclear factor-kB and of mitogen-activated protein kinases in macrophages. J Biol Chem 276:31906-31912 (¶on deputation/work done elsewhere).

5.     ¶Fiumara P, Snell V, Li Y, Mukhopadhyay A, Younes M, Gillenwater AM, Cabanillas F, Aggarwal, B.B. and Younes. A (2001) Functional expression of RANK in Hodgkin’s disease cell lines. Blood 98:2784-2790 (¶on deputation/work done elsewhere).