along with3D printingIn the important position of medical biological research, scientists have created new biological3D printingInk, the development of progressive microfluidic technology, etc. Recently, the University of Glasgow has developed bacteria-loaded microgels as an autonomous 3D environment for stem cell engineering. This research has developed a one-step microfluidic system that can encapsulate stem cells and genetically engineered non-pathogenic bacteria in the so-called 3D pearl lace-like alginate. The salt microgel has a high level of monodispersity and cell viability.
In research, although most technologies rely on droplet extrusion, researchers are creating more efficient systems through a one-step droplet microfluidics approach. The preparation of pearlescent band microgels takes place at physiological pH, without any sheath material, channel size and overall design means to avoid shear stress on the cells and promote viability.
The prepared gel construct is unique in that it has spacer units as in a single microcapsule and the connectivity found in fiber constructs. Also, the spacing between the microcells and the connections connecting them are height-adjustable, resulting in a highly monodispersed pearl-lace interconnection structure. Related to the manufacturing process, pearl units benefit from slower cross-linking in addition to suppressed shear stress compared to interconnected units. This technology allows the manufacture of zoned but linked cell-laden hydrogels with unprecedented precision and control, which has been used as a low-cost 3D bioprinting prototype.
3D printingTechnology provides bacteria-loaded microgels for stem cell engineering” alt=”Biology for scientists3D printingTechnology provides bacteria-loaded microgels for stem cell engineering” />
(Microfluidic settings based on droplets)
(A) Schematic diagram of the microfluidic device and encapsulation of prokaryotic and eukaryotic cells.
(B) Image of capillary-based microfluidic device.
(C) Snapshot of pearl formation in the microfluidic device, where the parameters used to quantify the assembled pearl are indicated.
(D) Thread thickness graph with corresponding flow velocity (Y axis: water flow; X axis: oil flow).
(E) Thread thickness diagram.
(F) Map of the Pearl District.
For this project, the researchers created an in vitro 3D model to study the relationship between eukaryotes (marrow mesenchymal stem cells, hBM-MSC) and prokaryotic cells (engineered non-pathogenic bacteria Lactococcus lactis, Lactococcus lactis) The symbiosis of symbiosis.
Although bacteria are often used as affordable “producing organisms” for proteins in bioprinting, they can also serve as mechanisms to direct cell growth and differentiation. Glasgow researchers also used the bacteriostatic antibiotic sulfamethoxazole to prevent the growth of harmful bacteria.
Made four3D printingThe shapes, including straight lines, triangles, squares and circles, are arranged as follows:
Line-two discs (180 degree angle)
Triangle (60 degree internal angle)
Square four corners (internal angle 90 degrees)
Eight are round (135 degree internal angle, octagonal)
Microfluidic systems enable researchers to create “monodisperse” constructs suitable for applications such as drug screening, biological research, and personalized medicine.
3D printingTechnology provides bacteria-loaded microgels for stem cell engineering” alt=”Biology for scientists3D printingTechnology provides bacteria-loaded microgels for stem cell engineering” />
(SEM image of alginate construct with cells, phase difference image of alginate microgel and MSC in basal medium)
(A); Sodium alginate microgel and MSC in osteogenic medium
(B); Contains two Lactococcus lactis
(C) Alginate microgel of MSC of the colony, which expresses FNIII 7-10 or BMP-2 in a constitutive manner. The samples were fixed after two weeks of incubation. Scale bar: 100μm.SEM image of alginate constructed with MSC in basal medium, the image shows the label applied by the cells on the cross section of the alginate construct
(D); Two colonies of the alginate microgel containing MSC and Lactococcus lactis exceed the space
(E); Alginate microgel, MSC in osteogenic medium, round solid covering cells, cavities and thin membrane-like constructs
(F).Compared with their state in an aqueous medium, the hydrogels are slightly dehydrated/shrinked
The connectivity of pearl lace hydrogels can provide a gradient research method in which the population of each cell type can control its relative density. It can also be used for time series indexing studies and provide an average value for the low-cost, easy-to-manufacture 3D bio-printing prototypes demonstrated in this study.
The microgel in this study has been used as a proof of concept for modeling the adjustable platform, where the hydrogel as ECM and the production and release of growth factors can be designed at low cost and have high-precision spatio-temporal control. It has been trying to further design more aspects of the in vitro system, pave the way for cell research, and exercise greater control over the interaction with the adjustable dynamic ECM-like environment.
As the progress of bioprinting continues to dominate global research, scientists have created new bioprinting inks,3D printingMicro-surface, progressive microfluidic technology, etc.
3D printingTechnology provides bacteria-loaded microgels for stem cell engineering” alt=”Biology for scientists3D printingTechnology provides bacteria-loaded microgels for stem cell engineering” />
(Monodispersity, encapsulation and vitality of microbeads)
(A) Under laminar flow conditions, using a flow rate of 500 μl h-1 and a flow rate of 5000 μl h-1 for the external phase, the bead size (long axis) distribution of the resulting hydrogel in two miscible fluid streams. Analyze N≥5-10 microgels for each condition. The average length of the pearls formed was 167μm, and the RSD was 3.2%. Scale bar: 100μm.
(B) Encapsulation efficiency of cells (MSC and Lactococcus lactis) by the formed hydrogel. The cell count at each time point is the result of 8 measurements taken sequentially at 30-minute intervals at room temperature for 2 hours.
(C) Fluorescence image of a two-week-old alginate hydrogel with Lactococcus lactis and MSC. For MSC, the hydrogel was stained with the BacLight bacterial viability kit and viability/cytotoxicity kit of Lactococcus lactis. Both kits stain live cells in green (SYTO 9 and Calcein AM) and non-viable cells in red (propidium iodide and Ethidium homodimer-1). The 50:50 mixture of the kit is used for co-cultivation. Scale bar: 100μm.
(D) Viability diagram of Lactococcus lactis, MSC and co-culture.
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