Polyethylene glycol‐based cationically charged hydrogel beads as a new microcarrier for cell culture

Abstract

A new polyethylene glycol (PEG) based microcarrier was designed and examined by the attachment and growth of mouse fibroblast cells. In the design of microcarrier, a PEG-based macromonomer, polyethyleneglycol methacrylate (PEGMA), was selected as the main component of hydrogel beads since PEG is known as a nontoxic and biocompatible material. A relatively new cationic comonomer, N-[3-(dimethylamino)propyl]methacrylamide (DMAPM), with higher ionization ability with respect to the similar comonomers was used for providing cationic charge to the hydrogel structure. In the first part, a suspension copolymerization method was developed for the production of cationically charged hydrogel beads as a potential microcarrier for cell culturing. The suspension copolymerization by using ethylene dimethacrylate (EDM) as cross-linking agent and cyclohexanol as the diluent provided spherical, polydisperse poly(PEGMA-DMAPM-EDM) hydrogel beads with an average size of 121 microm. The hydrogel beads exhibited a pH-dependent swelling behavior. The L929 mouse fibroblast cells were cultured on poly(PEGMA-DMAPM-EDM) hydrogel beads with an initial concentration of 200,000 cells/mL. The cells were incubated in Dulbecco's modified Eagle's medium during 5 days and the cell proliferation was investigated at every 24 h. An effective cell attachment and growth up to 3.5 x 10(6) cells/mL were observed with the poly(PEGMA-DMAPM-EDM) hydrogel beads. The results indicated that the proposed microcarrier was a significant alternative to the hydrogel beads obtained by the copolymerization of 2-hydroxyethyl methacrylate and 2-dimethylaminoethylmethacrylate commonly used in microcarrier-facilitated cell culturing studies.

A Novel Shell-Structure Cell Microcarrier (SSCM) for Cell Transplantation and Bone Regeneration Medicine

“…The cell growth-induced microspherical conglutination (as shown in Fig. 9e and c), which has also been observed in other microcarrier models (10,12,27), is believed to be beneficial for the constitution of neo-tissues with better continuity and integrity. In this study, the cytocompatibility of the SSCMs with two different sizes (100~154 μm and 180~280 μm) was evaluated by cell loading test.…”

Partitioning microfluidic channels with hydrogel to construct tunable 3-D cellular microenvironments

“…The Reynolds numbers for an aqueous solution flowing at 15 μl/min in these microchannels were less than one in all cases (between 0.17 for the five-inlet device and 0.28 for the three-inlet device) and the resulting flows were laminar, as expected. [25] Among the large number of hydrogels proposed in the literature for cell culture, [7,8,[26][27][28] we chose to use the Growth Factor Reduced Matrigel matrix (MG) for four reasons: i) it is biologically-derived-it is a soluble extract of the basement membrane of murine tumoral epithelia, mainly composed of laminin and collagen IV; ii) it is commercially available, and has been used extensively both in vivo and in vitro in the culture of many types of cells; [29] iii) it is liquid at 4 ºC, but gels within 15 minutes at 22 ºC -37 ºC, and iv) it is sufficiently stiff after gelling [9] to form self-standing structures of gel. The main disadvantage of Matrigel is that because of its biological origin, it contains not only structural proteins, but also growth factors and enzymes.…”

“…Cells suspended in uncured hydrogel can also be plated on Petri dishes, and -after gelation-exposed to a solution of cytokine that covers the layer of gel. [7][8][9] Suspending cells in liquid hydrogel followed by curing makes it possible to co-culture different types of cells.…”

Chemically crosslinked alginate porous microcarriers modified with bioactive molecule for expansion of human hepatocellular carcinoma cells

“…At present, a wide variety of commercial microcarriers (e.g., Cytodex 1, DE‐52, Cytopore 1, etc.) with a diameter range 100–250 μm have been developed and applied to the large‐scale culture of fibroblast, keratinocytes, ostroblast, chondrocytes, hepatocyte, embryonic stem cell (ESC), mesenchymal stem cells (MSC), pluripotent stem cells, and so forth. In addition, various new microcarriers were applied for cell expansion.…”