A comparative gene-expression analysis of CD34+ hematopoietic stem and progenitor cells grown in static and stirred culture systems

Abstract

Static and stirred culture systems are widely used to expand hematopoietic cells, but differential culture performances are observed between these systems. We hypothesize that these differential culture outcomes are caused by the physiological responses of CD34 + hematopoietic stem and progenitor cells (HSPCs) to the different physical microenvironments created in these culture devices. To understand the genetic changes provoked by culture microenvironments, the gene expression profiling of CD34 + HSPCs grown in static and stirred culture systems was compared using SMART-PCR and cDNA arrays. The results revealed that 103 and 99 genes were significantly expressed in CD34 + cells from static and stirred systems, respectively. Of those, 91 have similar levels of expression, while 12 show differential transcription levels. These differentially expressed genes are mainly involved in anti-oxidation, DNA repair, apoptosis, and chemotactic activity. A quantitative molecular understanding of the influences of growth microenvironments on transcriptional events in CD34 + HSPCs should give new insights into optimizing culture strategies to produce hematopoietic cells.

Expansion and hepatic differentiation of rat multipotent adult progenitor cells in microcarrier suspension culture

“…The feasibility of cultivating different stem cells has been demonstrated for various types of bioreactors such as perfusion reactors, rotating vessels, spinner reactors, and others (Cameron et al, 2006;Fok and Zanstra, 2005;Gilbertson et al, 2006;Li et al, 2006;Liu et al, 2006;Zhao et al, 2005;Bauwens et al, 2005;Schroeder et al, 2005). Ideally, a bioreactor should provide a homogeneous environment, for a high concentration of cells, and allow easy access for sampling.…”

“…Employing a scalable reactor system can enable a sufficient supply of differentiation competent or differentiated cells to meet such a clinical need. Since the conventional method of expanding and differentiating stem cells in tissue culture plates is labor intensive and difficult to scale up, different culture systems for expansion and differentiation of stem cells have been investigated; rotary culture systems, perfusion culture systems, and stirred suspension systems have been employed for various stem cell cultures (Cameron et al, 2006;Fok and Zanstra, 2005;Gilbertson et al, 2006;Li et al, 2006;Liu et al, 2006;Zhao et al, 2005;Bauwens et al, 2005;Schroeder et al, 2005). Among those systems, stirred suspension culture has the advantage of having been widely used in growing mammalian cells for decades.…”

“…It also allows high-density culture in a more homogeneous environment, and should allow one to monitor and control culture parameters. Stirred reactor cultures have been used to expand or differentiate hematopoietic stem and progenitor cells (HSPCs), neural stem cells (NSCs), and human and mouse embryonic stem cells (ESCs) grown as embryoid bodies (EBs) (Cameron et al, 2006;Fok and Zanstra, 2005;Gilbertson et al, 2006;Li et al, 2006). For cells grown as monolayers like those on tissue culture plates, microcarriers have the advantage of providing a large surface area for a given reactor volume.…”

Bioreactor development for stem cell expansion and controlled differentiation

“…Hematopoietic stem and progenitor cells (HSPCs) were among the first stem cells to be cultured in bioreactors. Two recent studies have confirmed that HSPC expansion is more extensive in stirred or suspended culture [2,3]. In contrast to HSPCs, most bioreactor studies with mesenchymal stem cells (MSCs) have employed perfusion of cells embedded within a 3-dimensional (3-D) scaffold.…”

Hematopoietic reconstitution of CD34+ cells derived from short-term cultured cord blood mononuclear cells

“…The magnetic separation step was repeated to further purify CD34 + cells. The purity of isolated CD34 + cell population is stated as > 95% [14].…”