BIOPRINTING, SIMPLIFIED 3D bioprinting allows researchers to model disease, create custom 3D tissue, and study the body outside the body with a level of control and repeatability that was simply not possible as recently as 5 years ago. For pharmaceutical development, 3D bioprinting offers a means of testing drugs faster, at a lower cost, and with greater biological

ALLEVI 3 - ALLEVI

Temperature Control. With temperature control from 4° C to 160° C, the Allevi 3 bioprinter is able to print a wide range of bioinks including collagen solutions, matrigel solutions, gelatin methacrylate, alginate, graphene, carbohydrate glass, polycaprolactone (PCL), and PLGA – just to name a few. QUICKSTART GUIDE: ALLEVI 2 The world’s first desktop 3D bioprinter. The Allevi 2 offers visible or UV light photocuring, room temperature to 160°C heating, and dual extruders. Find your bioink of choice and start building. This quickstart guide for the Allevi 2 has manuals and videos to help you get started with your new Allevi 2 GUIDE TO PICKING YOUR BIOPRINTING NEEDLE Guide to Picking Your Bioprinting Needle. Updated on March 18, 2020. Choosing a needle tip is a key component of the print parameter optimization process. It has a significant effect on the optimal printing speed and pressure, as well as the resolution of the print. The needle also determines the shear stress that the cells experience,

which

GETTING STARTED WITH G-CODE As the intermediate between STL models and a bioprinted model, gcode is a scripting language, or set of instructions, for controlling a 3D printer or bioprinter. Gcode holds a lot of importance but often isn’t understood well. Being able to read g-code is the first step to troubleshooting your models more effectively and gaining a better understanding of your bioprinter. BIOPRINTING SPHEROIDS/ORGANOIDS USING CUSTOM G-CODE Methods: Bioprinting Spheroids/Organoids Using Custom G-code. Create your G-code file by using the following template: Note: Allevi 2 users may need to enter M106 T_ as well to turn the extruder on. Similarly, they may need to enter M107 T_ to turn the extruder off. Copy and

paste lines 3

PASSAGING CELLS (GENERAL) Following is an easy to follow, step-by-step guide to Passaging Cells (General), including required materials and equipment. PRINT PARAMETER OPTIMIZATION GUIDE Results: This protocol will help you determine the material’s print pressure. Figure 1. Fill a syringe with your material and load it into the Allevi. In the Allevi software, press and hold the extrude button. Release to stop extruding. If there was no extrusion or uneven extrusion, increase the pressure by 5 HOW TO PREPARE GELMA BIOINK (LYOPHILIZED) Example: If you wish to make 10 mL of GelMA bioink, add 5 mL of cell media. Place a stirrer in your vial and leave it on the hot plate at 60˚C until all LAP is dissolved (15-30 min); Note: Ensure that your container is tightly sealed to avoid evaporation. Aliquot the amount of GelMA needed to make a 20% w/v solution; Example: To make a 20% w/v PASSAGING HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS Following is an easy to follow, step-by-step guide to Passaging Human Umbilical Vein Endothelial Cells (HUVECs), including required materials and equipment. BIOPRINTING, SIMPLIFIED 3D bioprinting allows researchers to model disease, create custom 3D tissue, and study the body outside the body with a level of control and repeatability that was simply not possible as recently as 5 years ago. For pharmaceutical development, 3D bioprinting offers a means of testing drugs faster, at a lower cost, and with greater biological

ALLEVI 3 - ALLEVI

Temperature Control. With temperature control from 4° C to 160° C, the Allevi 3 bioprinter is able to print a wide range of bioinks including collagen solutions, matrigel solutions, gelatin methacrylate, alginate, graphene, carbohydrate glass, polycaprolactone (PCL), and PLGA – just to name a few. QUICKSTART GUIDE: ALLEVI 2 The world’s first desktop 3D bioprinter. The Allevi 2 offers visible or UV light photocuring, room temperature to 160°C heating, and dual extruders. Find your bioink of choice and start building. This quickstart guide for the Allevi 2 has manuals and videos to help you get started with your new Allevi 2 GUIDE TO PICKING YOUR BIOPRINTING NEEDLE Guide to Picking Your Bioprinting Needle. Updated on March 18, 2020. Choosing a needle tip is a key component of the print parameter optimization process. It has a significant effect on the optimal printing speed and pressure, as well as the resolution of the print. The needle also determines the shear stress that the cells experience,

which

GETTING STARTED WITH G-CODE As the intermediate between STL models and a bioprinted model, gcode is a scripting language, or set of instructions, for controlling a 3D printer or bioprinter. Gcode holds a lot of importance but often isn’t understood well. Being able to read g-code is the first step to troubleshooting your models more effectively and gaining a better understanding of your bioprinter. BIOPRINTING SPHEROIDS/ORGANOIDS USING CUSTOM G-CODE Methods: Bioprinting Spheroids/Organoids Using Custom G-code. Create your G-code file by using the following template: Note: Allevi 2 users may need to enter M106 T_ as well to turn the extruder on. Similarly, they may need to enter M107 T_ to turn the extruder off. Copy and

paste lines 3

PASSAGING CELLS (GENERAL) Following is an easy to follow, step-by-step guide to Passaging Cells (General), including required materials and equipment. PRINT PARAMETER OPTIMIZATION GUIDE Results: This protocol will help you determine the material’s print pressure. Figure 1. Fill a syringe with your material and load it into the Allevi. In the Allevi software, press and hold the extrude button. Release to stop extruding. If there was no extrusion or uneven extrusion, increase the pressure by 5 HOW TO PREPARE GELMA BIOINK (LYOPHILIZED) Example: If you wish to make 10 mL of GelMA bioink, add 5 mL of cell media. Place a stirrer in your vial and leave it on the hot plate at 60˚C until all LAP is dissolved (15-30 min); Note: Ensure that your container is tightly sealed to avoid evaporation. Aliquot the amount of GelMA needed to make a 20% w/v solution; Example: To make a 20% w/v PASSAGING HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS Following is an easy to follow, step-by-step guide to Passaging Human Umbilical Vein Endothelial Cells (HUVECs), including required materials and equipment.

ABOUT - ALLEVI

About Allevi. Allevi creates tools and solutions to design, engineer and build with life. Our 3D bioprinters and bioinks are used around the world to find solutions to humanity’s most difficult problems – to cure disease, to test novel drugs, and to eliminate the organ transplant waiting list. Founded in 2014, our mission is to make it

easy

THE FRESH METHOD

The FRESH method utilizes a support hydrogel as a temporary, thermoreversible support that can be washed away after printing. The FRESH support reagent is comprised of processed gelatin microparticles with a Bingham plastic rheology. This biocompatible material offers temporary support of fragile materials used for bioprinting. QUICKSTART GUIDE: ALLEVI 1 Small, friendly, and powerful. The Allevi 1 packs autocalibration, 4-160°C temperature control, and visible/UV photocuring into a small footprint. Find your bioink of choice and start building. Follow this quickstart guide of manuals and videos to help you get started with

your new Allevi 1

CELLS AND BIOINKS

Bioinks are natural or synthetic biomaterials that mimic the extracellular matrix (ECM) to support the adhesion, proliferation, and differentiation of living cells. These materials give cells important cues they need to live, grow, and create functional 3D tissue. GELATIN METHACRYLATE (GELMA) GUIDE Bioink Review: Gelatin methacrylate, commonly referred to as gelMA, is a photocrosslinkable natural bioink derived from a hydrolytic degradation of collagen (1).GelMA is commonly used in drug delivery systems and wound dressing applications (1). With its cell encapsulation properties and thixotropic nature, gelMA is also a common reagent used for bioprinting (8).

BIOPRINTING GELMA

Methods. Mix LAP in 3 ml of PBS or cell media at 60°C under stirring to create a 0.5% w/v LAP concentration. Mix until all LAP is dissolved. Add GelMA to solution to create a 10% w/v concentration. Mix at 60°C under stirring until all GelMA is dissolved. Be sure the container is tightly sealed while mixing to avoid evaporation of

solution.

BIOPRINTING PARAMETERS FOR EVERY BIOINK CHEAT SHEET Welcome to the Allevi quick start guide to bioprinting parameters for every bioink! The following bioprinting parameters are general guidelines for bioinks offered on the Allevi platform, but with some simple experiments, you can optimize them for your exact application!. Every bioink requires different parameters in order to achieve high printability and viability. INFILL PATTERN OPTIONS IN SLIC3R The infill was set to 10% and the filament diameter to 0.3mm to help illustrate the infill patterns. The top layer is removed in the images in order to show the inner pattern. Note: The circles were sliced with a concentric base. Note: The square were sliced with a rectilinear base. 3D Matter goes into a deeper discussion of infill percentage

GAUGE SIZE GUIDE

Gauge Size Guide. Needle gauges can be confusing. And as an important parameter for bioprinting, you want to select the correct tip for your syringes. The gauge size of a needle indicates what the diameter of the lumen (opening) is. The higher the gauge, the smaller the diameter. For example, a 30 gauge needle has a narrower lumen than a

25

HEMOCYTOMETER CELL COUNTING Use the following formula in order to calculate the number of cells you have in your suspension: (total cells counted)/(4 squares counted)*10-4 *initial volume*dilution factor = total number of cells; Note: 10-4 is the volume of squares on the hemocytometer (0.1 mm 3).The dilution factor will be 1, unless you have diluted your cell suspension with trypan blue.

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3D BIOPRINTING, SIMPLIFIED Bring your work to life with Allevi 3D bioprinters, materials, and

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ALLEVI 3 BIOPRINTER

The bioprinter for pharmaceutical development, tissue engineering, and everything in between. Meet the Allevi 3 →

COMPARE BIOPRINTERS

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ALLEVI 1

With the smallest footprint and widest material capabilities of any 3D bioprinter on the market, you’re going to love using the Allevi 1.

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ALLEVI 2

The Allevi 2 is the world’s first desktop 3D bioprinter. It is a beautifully designed, precision manufactured 3D bioprinter that prints

cells and bioinks.

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WHY 3D BIOPRINTING?

3D bioprinting allows researchers to model disease, create custom 3D tissue, and study the body outside the body with a level of control and repeatability that was simply not possible as recently as 5 years

ago.

For pharmaceutical development, 3D bioprinting offers a means of testing drugs faster, at a lower cost, and with greater biological relevance than animal testing or 2D _in vitro_ studies. In the biomedical devices field, 3D bioprinting has enabled new developments such as sugar stents to help surgeons join veins

with

fewer complications, and

systems

for improved drug delivery

.

Personalized and regenerative medicine continue to grow in popularity, and bioprinting will give doctors and researchers the tools to better target treatments and improve patient outcomes. Over 110,000 people in the US

alone

are on the organ transplant waiting list at any given time. There’s a large and growing need for an alternative to organ and tissue donation. 3D bioprinting is an exciting new tool with the potential to eliminate the organ transplant waiting list More About Bioprinting →

FROM THE BLOG

Bioprinting with Corning Matrigel Matrix After 30 years of discoveries and over 10,000 citations, Corning® Matrigel® matrix has cemented its place as a trusted ECM in labs worldwide. Biologists, tissue engineers, and industry giants alike rely on Matrigel matrix as a foundational biomaterial for diverse applications;... Allevi Designs for Different Futures We've always thought the Allevi 1 was a work of art, but now it's official! We're honored to be included in a major exhibition on display at the Philadelphia Museum...

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Allevi Author: How 3D Bioprinting Solves the Stem Cell Supply Problem If stem cell therapies take off - we're going to need a way to solve the stem cell supply problem - read on to learn more: The Promise

We’ve all...

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Bioprinting with Corning Matrigel Matrix After 30 years of discoveries and over 10,000 citations, Corning® Matrigel® matrix has cemented its place as a trusted ECM

in...

Read More__

Allevi Designs for Different Futures We've always thought the Allevi 1 was a work of art, but now it's official! We're honored to be included...

Read More__

Allevi Author: How 3D Bioprinting Solves the Stem Cell Supply Problem If stem cell therapies take off - we're going to need a way to solve the stem cell supply problem...

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