Any help is greatly appreciated. I want to learn about this particular aspect of the tissue engineering process.
Now， I just do the experiments about the cell culture in alginate sodium, those are the related references, you can have a look
1.Mass Production of Cell-Laden Calcium Alginate Particles
with Centrifugal Force
2.Microfluidic Templated Multicompartment Microgels for
3D Encapsulation and Pairing of Single Cells
3.Compartmentalized Spherical Collagen Microparticles for
Anisotropic Cell Culture Microenvironments
Hi Alberta Kovatcheva
The gelation of alginate is most commonly performed through crosslinking polymers with divalent cations, such as calcium.
This simple method of gelation allows the formation of alginate hydrogels of different shapes and dimensions, but most frequently alginate beads are produced.
The method for bead generation can be performed by either external or internal gelation.
Good luck !
CELL ENCAPSULATION
- Harvest cells from monolayer cultures using trypsin-EDTA.
- Filter them through a 40 μm pore mesh.
- Suspend cells in a solution of 1.5% (w/v) (variable) sodium alginate (for example, UPLVG from FMC Biopolymer, Norway) in phosphate buffered saline (PBS, w/o Ca++), at desired cell density (typically, 5 million/mL).
- Switch on droplet generator (electrostatic, air flow, vibrational,…) (this gives for another thread).
- Extrude cell suspension in a sterile syringe through a 0.35 mm needle (variable) at a 5.9 mL/h flow rate (variable) using a peristaltic pump.
- Collect the resulting alginate particles in a 55 mM CaCl2 (variable) solution and maintain them under agitation for 15 min after the end of the process to ensure complete gelation of all the beads.
- Wash with DPBS w/Ca++.
OPTIONAL COATINGS
- Suspend the beads in 0.05% PLL/PLO (or other polycation) solution for 5 min (variable)
- Wash with DPBS w/Ca++.
- Coat again with another layer of 0.1% alginate for 5 min (variable).
- Wash twice with DPBS w/Ca++.
For all the solutions pH and osmolarity values must be adjusted to 7.4 (HEPES buffer) and 270 mOsm/L, respectively.
All the process must be carried out under aseptic conditions, at room temperature.
Resulting microcapsules may be cultured in complete medium at 37 °C in a 5% C02/95% air atmosphere standard incubator.
Hi Alberta,
Recently, I completed some work in encapsulation of MSCs in sodium alginate beads. Although, many articles reported very good methods about encapsulation, as other people here also gave very good & explanatory answers. I followed few papers which gave me overall idea of encapsulation work. Hope you also might have found the similar papers. still please find attached links.
Article Encapsulation of Mesenchymal Stem Cells from Wharton's Jelly...
Article Alginate microcapsule as a 3D platform for the efficient dif...
Article Alginate microcapsule for propagation and directed different...
Article Osteogenic Differentiation of Human Mesenchymal Stem Cells i...
Article Dental pulp stem cells immobilized in alginate microspheres ...
Saurabh
Here is a protocol for encapsulation that has been used for stem cells. Full text available on research gate. best, brad
https://www.nature.com/articles/nprot.2011.352
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