Engineering a Human Bone Marrow Model: A Case Study on ex Vivo Erythropoiesis

Abstract

Bone marrow, with its intricate, three-dimensional tissue structure facilitating cell-cell interactions, provides a microenvironment supporting the production of hundreds of billions of multilineal blood cells everyday. We have developed a three-dimensional bone marrow culture system in which marrow cells are cultured in a reactor packed with porous microspheres. The culture supports a three-dimensional growth configuration and multilineal hemopoiesis mimicking the bone marrow in vivo. We studied ex vivo human erythropoiesis using the three-dimensional culture system. The system sustained extensive erythropoiesis at low erythropoietin concentrations (0.2 U/mL), plus stem cell factor, interleukin-3, granulocyte-macrophage colony-stimulating factor, and insulin-like growth factor-I. Erythroid cell production lasted for more than 5 weeks, and the percentage of erythroid cells in the nonadherent cell population was approximately 60%. Flow cytometric analysis using cell surface markers specific for erythroid cells (CD71 and glycophorin-A) indicated that the culture produced early, intermediate, and late erythroid cells. As the culture progressed, the erythroid cell population shifted gradually toward mature cell types. When compared to the three-dimensional culture, the traditional flask cultures failed to support extensive erythropoiesis under the same conditions. This indicates that the three-dimensional bone marrow culture system provides a microenvironment conducive to erythropoiesis under more physiological conditions and is a better bone marrow model.

Development and molecular characterization of polymeric micro-nanofibrous scaffold of a defined 3-D niche for in vitro chemosensitivity analysis against acute myeloid leukemia cells

“…Despite many attempts toward the recreation of an in vivo environment closer to the bone marrow niche in vitro, the success rate has been poor. 4,12,[32][33][34][35] Since the bone marrow niche is a hideout for LSCs, it is pertinent to generate a 3-D environment for drug testing and perhaps ex vivo production of blood cells. [5][6][7][8][9][10][11][12] Although there have been attempts toward the fabrication of scaffolds that resemble the bone marrow niche with better cell adhesion and proliferation compared to 2-D matrices, the development of a micro-nanofibrous environment close to the bone marrow niche, without any added stromal cells, showing similar in vivo molecular characteristics to CAMDR has not yet been shown, as far as we are aware.…”

Large‐scale production of red blood cells from stem cells: What are the technical challenges ahead?

“…Nonetheless, many attempts have been made to scale up HSC cultures over the past twenty years, aiming at increasing the number of useful stem cells obtained from limited quantities collected, for cell therapy and transplantation following chemotherapy. A wide range of bioreactors have been tested: airlift bioreactors [71], spinner flasks [69,71,[88][89][90][91][92] and stirred bioreactors [93,94], hollow-fibers [71], fixed-bed reactors with microcarriers [91,95,96] and rotating wall vessel reactors [97,98]. The data have been compared [76,98], with best results (total cell expansion up to 435-fold, 32-fold expansion of CD34 + cells and 21-fold expansion of colony forming unit-granulocyte/macrophages (CFU-GM)) for stirred and rotating wall vessel reactors, far beyond the yields in static flasks.…”

Two-Dimensional Polymer-Based Cultures Expand Cord Blood-Derived Hematopoietic Stem Cells and Support Engraftment of NSG Mice

“…(C) Representative human CD45 + , CD34 + , CD13 + , and CD19 + engraftment in the BM of NSG transplanted mice. Color images available online at www.liebertpub.com/tec Up to now, only a few systems for HSC culture using biomaterials have been proposed, including the use of collagen, 17,19,29,30 nylon meshes, 31 tantalum-coated carbon fibers, 32 PET matrices, 15,16,18 polyethylene-alt-maleic anhydride, 33 PLGA, and PU. 20 However, biomaterial-based studies in general resulted in highly variable outcomes, mainly due to concerns such as: (1) the choice of using biomaterial types is nearly arbitrary and (2) the effect of using different culture media in combination with biomaterials is itself not well characterized.…”