Effect of Growth Factors on Ex Vivo Bone Marrow Cell Expansion Using Three‐Dimensional Matrix Support

Tun, Thein; Miyoshi, Hirotoshi; Aung, Tin; Takahashi, Satoru; Shimizu, Ritsuko; Kuroha, Takashi; Yamamoto, Masayuki; Ohshima, Norio

doi:10.1046/j.1525-1594.2002.06842.x

Cited by 34 publications

(32 citation statements)

References 28 publications

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“…3D systems have been employed to culture HSCs in recent years. Many scaffolds have been used, which included natural materials, such as collagen carriers and cellulose porous microspheres, or synthetic materials, such as porous biomatrix, polyethylene terephthalate, porous polyvinyl formal, porous gelatin microspheres, polyester nonwoven fabric porous disc carriers, and colloidal crystals (Bagley et al 1999;Li et al 2001;Zhang et al 2006;Tun et al 2002;Tomimori et al 2000;Nichols et al 2009). These culture systems were superior to traditional 2D systems for maintaining and expanding the HSCs/HPCs.…”

Section: Discussionmentioning

confidence: 99%

Maintenance and expansion of hematopoietic stem/progenitor cells in biomimetic osteoblast niche

Jinquan

Liu

et al. 2010

Cytotechnology

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In this study, we employed bio-derived bone scaffold and composited with the marrow mesenchymal stem cell induced into osteoblast to replicate a "biomimetic niche." The CD34(+) cells or mononuclear cells (MNC) from umbilical cord blood were cultured for 2-5 weeks in the biomimetic niche (3D system) was compared with conventional two dimensional cultures (2D system) without adding cytokine supplement. After 2 weeks in culture, the CD34(+) cells from umbilical cord blood in the 3D system increased 3.3-4.8 folds when compared with the initial CD34(+) cells. CD34(+)/CD38(-) cells accounted for 82-90% of CD34(+) cells. After 5 weeks, CD34(+)/CD38(-) cells in the 3D system increased when compared with initial (1.3 ± 0.3 × 10(3) vs. 1.0 ± 0.5 × 10(4), p < 0.05), but were decreased in the 2D system (1.3 ± 0.3 × 10(3) vs. 2.5 ± 0.7 × 10(2), p < 0.05). The CFU progenitors were produced more in the 3D system than in the 2D system (4.6-9.3 folds vs. 1.0-1.5 folds) after 2 weeks in culture, and the colony distribution in the 3D system manifested higher percentage of BFU-E and CFU-GEMM, but in the 2D system was mainly CFU-GM. The LTC-ICs in the 3D system showed 5.2-7.2 folds increase over input at 2 weeks in culture, and maintain the immaturation of hematopoietic progenitor cells (HPCs) over 5 weeks. In conclusion, this new 3D hematopoietic progenitor cell culture system is the first to utilize natural cancellous bone as scaffold with osteoblasts as supporting cells; it is mimicry of natural bone marrow HSC niche. Our primary work has demonstrated it could maintain and expand HSC/HPC in vitro.

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Section: Discussionmentioning

confidence: 99%

Maintenance and expansion of hematopoietic stem/progenitor cells in biomimetic osteoblast niche

Jinquan

Liu

et al. 2010

Cytotechnology

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“…Considering the in vivo scenario, one realizes that each organ constitutes the functional cell-mass within extra cellular matrices in a three dimensional configuration. Researchers have used natural/artificial matrices to design three dimensional microenvironment for maintenance of tissue architecture in vitro viz collagen I matrix to generate skin organotypic cultures (Gron et al 1999), porous titanium fiber mesh to study osteoblast differentiation (Sikavitsas et al 2003), Calcium cross linked alginate to study marrow cell proliferation (Wang et al 2003) and porous polyvinyl formal (PVF) resin scaffold to study ex vivo expansion of human progenitor cells (Tun et al 2002). All these data confirm the superiority of three-dimensional growth conditions over two dimensional growth.…”

Section: Suitability Of Hanging Drop Technique For Cytotoxicity Studiesmentioning

confidence: 89%

Application of hanging drop technique for stem cell differentiation and cytotoxicity studies

Banerjee

Bhonde

2006

Cytotechnology

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The aim of our study is to explore the possibility of using an ancient method of culture technique- the hanging drop technique for stem cell differentiation and cytotoxicity testing. We demonstrate here a variety of novel applications of this age old technique not only to harness the differentiation potential of stem cells into specific lineages but also for cytotoxicity studies. Here we have prepared hanging drop cultures by placing 20 microl micro-drops of nutrient media and 10% Fetal Calf Serum (FCS) containing cells of interest on the lids of 60 mm dishes. Bottom plates of the dishes were filled with sterile Phosphate Buffer Saline (PBS) to avoid desiccation of samples. Lids were then placed on the bottom plates to achieve hanging drop cultures. We utilized this technique for cultivation of ciliated epithelia to study cytotoxicity and differentiation of bone marrow stromal cells. Most importantly the modified culture technique presented here is simple, economical and cost effective in terms of the time taken and the reagents required and are amenable to goal specific modification such as cytotoxicity testing. It is advantageous over the existing system in terms of retention of viability and functionality for longer duration and for providing three dimensional growth micro-environment making it useful for organotypic cultures and in vivo simulation.

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“…Much attention has been given in previous years to the design of matrices able to promote the appropriate spatial cell arrangement. In addition to the obvious advantage of a high surface area per volume ratio compared to monolayer systems, culturing in 3D matrices also enhances cell-cell and cell-matrix interactions and allows better cell distribution [48]. Specifically, the cultivation of hESC in scaffolds was shown to result in an appropriate cell differentiation and neo-tissue formation, including blood vessels, and an integration with the host upon implantation [49], in contrast to the incomplete differentiation observed in two-dimensional (2D) studies.…”

Section: Synthetic Extracellular Matrixmentioning

confidence: 99%

Microenvironment Design for Stem Cell Fate Determination

Ra'em

Cohen

2011

Tissue Engineering III: Cell - Surface Interactions for Tissue Culture

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Stem cells are characterized by their dual ability for self-renewal and differentiation, potentially yielding large numbers of cells that can be used in cell therapy and tissue engineering for repairing devastating diseases. Attaining control over stem cell fate decision in culture is a great challenge since these cells integrate a complex array of “niche” signals, which regulate their fate. Given this, the recent findings that synthetic microenvironments can be designed to gain some level of control over stem cell fate are encouraging. This chapter provides an overview of the current state and knowledge of the design of synthetic microenvironments bio-inspired by the adult stem cell niche. We describe the biomaterials used for reconstituting the niche, highlighting the bioengineering principles used in the process. Such synthetic microenvironments constitute powerful tools for elucidating stem cell regulatory mechanisms that should fuel the development of advanced culture systems with accurate regulation of stem cell fate.

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Effect of Growth Factors on Ex Vivo Bone Marrow Cell Expansion Using Three‐Dimensional Matrix Support

Cited by 34 publications

References 28 publications

Maintenance and expansion of hematopoietic stem/progenitor cells in biomimetic osteoblast niche

Maintenance and expansion of hematopoietic stem/progenitor cells in biomimetic osteoblast niche

Application of hanging drop technique for stem cell differentiation and cytotoxicity studies

Microenvironment Design for Stem Cell Fate Determination

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