Cultivation of Hematopoietic Stem and Progenitor Cells: Biochemical Engineering Aspects

Noll, Thomas; Jelinek, Nanni; Schmidt, Sebastian; Biselli, M.; Wandrey, Christian

doi:10.1007/3-540-45736-4_6

Cited by 22 publications

(15 citation statements)

References 80 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Despite extensive research conducted over the past 20 or more years, HSC in vitro culture expansion and differentiation toward blood cell lineages is not sufficiently optimized for industrial production 3 . In general, there are many difficulties and process challenges relating to HSC cultivation for the production of type O, Rhnegative RBCs in standard Petri dish cultures and in bioreactor systems 4,5 . Such challenges can be attributed in part to difficulties encountered during culture, which include but are not limited to donor variability 6 , recreation of a complex microenvironment 7 , and the interaction between various culture parameters 8 .…”

Section: Introductionmentioning

confidence: 99%

Compartmental Hollow Fiber Capillary Membrane–Based Bioreactor Technology for In Vitro Studies on Red Blood Cell Lineage Direction of Hematopoietic Stem Cells

Housler

Miki

Schmelzer

et al. 2012

Tissue Engineering Part C: Methods

View full text Add to dashboard Cite

Continuous production of red blood cells (RBCs) in an automated closed culture system using hematopoietic stem cell (HSC) progenitor cell populations is of interest for clinical application because of the high demand for blood transfusions. Previously, we introduced a four-compartment bioreactor that consisted of two bundles of hollow fiber microfiltration membranes for transport of culture medium (forming two medium compartments), interwoven with one bundle of hollow fiber membranes for transport of oxygen (O 2 ), carbon dioxide (CO 2 ), and other gases (forming one gas compartment). Small-scale prototypes were developed of the three-dimensional (3D) perfusion cell culture systems, which enable convection-based mass transfer and integral oxygenation in the cell compartment. CD34+ HSC were isolated from human cord blood units using a magnetic separation procedure. Cells were inoculated into 2-or 8-mL scaled-down versions of the previously designed 800-mL cell compartment devices and perfused with erythrocyte proliferation and differentiation medium. First, using the small-scale 2-mL analytical scale bioreactor, with an initial seeding density of 800,000 cells/mL, we demonstrated approximately 100-fold cell expansion and differentiation after 7 days of culture. An 8-mL laboratoryscale bioreactor was then used to show pseudocontinuous production by intermediately harvesting cells. Subsequently, we were able to use a model to demonstrate semicontinuous production with up to 14,288-fold expansion using seeding densities of 800,000 cells/mL. The down-scaled culture technology allows for expansion of CD34 + cells and stimulating these progenitors towards RBC lineage, expressing approximately 40% CD235 + and enucleation. The 3D perfusion technology provides an innovative tool for studies on RBC production, which is scalable.

show abstract

Section: Introductionmentioning

confidence: 99%

Compartmental Hollow Fiber Capillary Membrane–Based Bioreactor Technology for In Vitro Studies on Red Blood Cell Lineage Direction of Hematopoietic Stem Cells

Housler

Miki

Schmelzer

et al. 2012

Tissue Engineering Part C: Methods

View full text Add to dashboard Cite

show abstract

“…Numerous studies have demonstrated that various cultivation conditions, including the cytokines cocktails, oxygen tension, pH, culture media and osmolality, may alter the expansion effects of human stem and progenitor cells (HSPCs) (Noll et al, 2002). Generally, HSPCs expanded ex vivo cannot be used in clinics due to their biological incompatibility.…”

Section: Introductionmentioning

confidence: 99%

Role of the plasma membrane ROS-generating NADPH oxidase in CD34+ progenitor cells preservation by hypoxia

Fan

Cai

Tan

2007

Journal of Biotechnology

View full text Add to dashboard Cite

Hypoxia favored the preservation of progenitor characteristics of hematopoietic stem and progenitor cells (HSPCs) in bone marrow. This work aimed at studying the role of reactive oxygen species (ROS)-generating NADPH oxidase system regulated by hypoxia in ex vivo cultures of cord blood CD34 + cells. The results showed that NADPH oxidase activity and ROS generation were reduced in hypoxia with respect to normal oxygen tension. Meanwhile the ROS generation was found to be inhibited by diphenyleneiodonium (the NADPH oxidase inhibitor), or N-acetylcysteine (the ROS scavenger). Accordingly NADPH oxidase mRNA and p67 protein levels decreased in hypoxia. The analysis of progenitor characteristics, including the proportion of cultured cells expressing the HSPCs marker CD34 + CD38 − , colony production ability of the colony-forming cells (CFCs), and the re-expansion capability of the cultured CD34 + cells, showed that either 5% pO 2 or reduced ROS favored preserving the characteristics of CD34 + progenitors, and promoted the expansion of CD34 + CD38 − cells as well. The above results demonstrated that hypoxia effectively maintained biological characteristics of CD34 + cells through keeping lower intracellular ROS levels by regulating NADPH oxidase.

show abstract

“…Thus, how to create an effective hematopoietic environment in vitro, and expand HSCs with normal biological activity is an urgent task for researchers. So far, researches have been focusing more on the positive regulation of HSCs, such as cytokines combination, the co-culture with stroma cells and hypoxia etc [3] , but less attention has been paid to negative regulators. Recently, ROS and p38MAPKα were demonstrated to be involved in the maintenance of HSCs quiescence [4][5][6] .…”

mentioning

confidence: 99%

The cross-talk between ROS and p38MAPKα in the Ex Vivo expanded human umbilical cord blood CD133+ cells

Zou

Lou

et al. 2011

J. Huazhong Univ. Sci. Technol. [Med. Sci.]

View full text Add to dashboard Cite

This study investigated the correlation between and compared the effects of reactive oxygen species (ROS) and p38 mitogen-activated protein kinase α (p38MAPKα) in the ex vivo expanded umbilical cord blood (hUCB) CD133(+) cells. hUCB CD133(+) cells were cultured in the hematopoietic stem cells (HSCs) culture medium with N-acetylcysteine (NAC, an anti-oxidant), p38MAPKα-specific inhibitor (SB203580) or their combination. The levels of ROS and expression of phosphorylated p38MAPKα (p-p38) in CD133(+) cells were flow cytometrically detected. The efficacy of ex vivo expansion was evaluated by the density of CD133(+) cell sub-group colony-forming cells (CFC) and cobblestone area-forming cells (CAFC) assay. Our results showed decreased ROS levels in NAC, SB203580, and their combination treatment groups were almost 37%, 48%, and 85%, respectively. Furthermore, SB203580 abrogated the activation of p38MAPKα more obviously than NAC. Moreover, the CD133(+) cells in SB203580 treatment group had a 21.93±1.36-fold increase, and 14.50±1.19-fold increase in NAC treatment group, but only 10.13±0.57-fold increase in control group. In addition, SB203580 treatment led a higher level increase in the number of CFU and CAFC than NAC did. These findings suggested that, in expanded CD133(+) cells, ROS activates p38MAPKα, which, in turn, induces ROS production, and p38MAPKα might be the most suitable regulator in ROS-p38MAPKα pathway for the promotion of HSCs ex vivo expansion.

show abstract

Cultivation of Hematopoietic Stem and Progenitor Cells: Biochemical Engineering Aspects

Cited by 22 publications

References 80 publications

Compartmental Hollow Fiber Capillary Membrane–Based Bioreactor Technology for In Vitro Studies on Red Blood Cell Lineage Direction of Hematopoietic Stem Cells

Compartmental Hollow Fiber Capillary Membrane–Based Bioreactor Technology for In Vitro Studies on Red Blood Cell Lineage Direction of Hematopoietic Stem Cells

Role of the plasma membrane ROS-generating NADPH oxidase in CD34+ progenitor cells preservation by hypoxia

The cross-talk between ROS and p38MAPKα in the Ex Vivo expanded human umbilical cord blood CD133+ cells

Contact Info

Product

Resources

About