Bio 3D Printer Regenova with Kenzan method

We provide the leading edge platform of three-dimensional organ regeneration using our revolutionary bio 3D printer, Regenova. Regenova is a novel robotic system that facilitates the fabrication of three- dimensional cellular structures by placing cellular spheroids in fine needle arrays, “Kenzan method”, according to pre-designed 3D data.

Bio 3D printer (Bioprinting) Regenova with Kenzan method

Platform Technology – 3D Bioprinting

Generation of thick and elastic tissue that is like real tissue but grown from cultured cells has proved difficult with conventional technologies.
Dr. Koichi Nakayama focused on the natural and intrinsic features of cell self-aggregation and invented a novel method to assemble cellular aggregates (spheroids) into any desired three-dimensional macroscopic tissue without collagen or hydrogel materials.
3D Bioprinting materials
With this method, cellular aggregates called spheroids are cultured, with several tens of thousands of cells per spheroid.
Then, spheroids are placed in fine needle arrays and allowed to merge with adjacent spheroids to form a connected structure that is firm enough not to require needles.
With appropriate alignment of needles to form the needle array, cellular spheroids can be positioned in any desired three-dimensional layout.
Finally, culturing connected cellular spheroids in a bioreactor promotes self-organization of cells and yields a 3D tissue with the desired function and quality.

The benefit of placing cellular spheroids in the needle array is not only freedom of three-dimensional positioning but also easier circulation of culture medium around the assembled spheroids to supply efficient neutralization and oxygen to cells until tissue is matured and no longer requires the support array.

Sphere-like cell

Sphere-like cell aggregates are secured in place by skewers.

The aggregates

The aggregates are allowed to fuse on the skewers for a few days.

The skewers

The skewers are removed and the tissue is allowed to further mature.

Proprietary Technology
The Kenzan method was invented by prof. Koich Nakayama (Saga University)and is globally patented intellectual property. Cyfuse Biomedical K.K. is granted exclusive rights of use.
3D Bioprinting process
STEP1 Loading spheroids and preparing the 3D data and needle array

Isolate cells and proliferate them to target levels. In low adhesion multi-well plates, several tens of thousands of cells form a spheroid in a day or two with a diameter of approximately 500 micrometers.

Spheroids photo

3D data photo

An example for low adhesion multi-well plates: PrimeSurface®96U (MS-9096UZ, S-BIO, Sumitomo Bakelite Co., Ltd.)

3D data

Design a three-dimensional arrangement of cellular spheroids and prepare 3D data using the original software. Multiple types of spheroids can be selected as desired.

3D data photo

To design three dimensional structure, we offer special software called “B3D”, which was developed for Regenova.
You can install B3D into your own computer and design no matter where you are.


Needle array

Needles are aligned to serve as a temporary scaffold during cellular spheroid assembly. The needles are made of stainless steel with a diameter of 100-200 micrometers and pitch of 300-400 micrometers.

Needle array photo

Needle array photo
STEP2 3D printing
3D printing photos
Regenova assembles cellular spheroids into a three-dimensional shape by placing spheroids in needle arrays according to the pre-designed 3D data.
STEP3 Maturation
Maturation photo
Further culturing of 3D printed tissue with proper flow of medium in a bioreactor facilitates rearrangement of cells for self-organization to exhibit physical strength and tissue function. The 3D shape is self-sustained without needles.

< An example to make 1cm^2 sheet with Mesenchymal Stem Cells >

3D printing photos

Obtained strong and elastic 1cm^2 sheet

Proprietary Technology
The Kenzan method was invented by prof. Koich Nakayama (Saga University)and is globally patented intellectual property. Cyfuse Biomedical K.K. is granted exclusive rights of use.

Manufacturing and distribution
Distributor: Cyfuse Biomedical K.K., Tokyo, Japan
Manufacturer: Shibuya Kogyo Co., Ltd., Ishikawa, Japan

Consumables and peripheral equipment
Needle array: Customers can select the proper shape and size of needle arrays depending on the tissue to be manufactured.
Bioreactor: A bioreactor is needed for efficient delivery of oxygen and nutrition to the solid tissue during the maturation step. Our scientists can consult regarding which system is suitable for the tissue of interest.

Bio 3D Printer Regenova, News and Events

Dec, 16, 2015 | 2015 cell biology ascb annual meeting on December 12 to 16, 2015

Dec, 10, 2015 | World Stem Cell Summit 15 on December 10 to 12, 2015

Nov, 09, 2015 | 3rd FAST Annual Congress (Functional Analysis & Screening Technologies) on November 9 to 11, 2015

Nov, 07, 2015 | AHA Scientific Sessions 2015 (American Heart Association) on November 7 to 11, 2015

Oct, 17, 2015 | Neuroscience 2015 on Oct 17 to 21, 2015

Sep, 17, 2015 | NIH Research Festival Exhibit on 17th – 18th September, 2015

Sep, 08, 2015 | 4th TERMIS World Congress (Tissue Engineering International & Regenerative Medicine Society) on September 8 to 11, 2015

Jul, 27, 2015 | 3D design software called B3D is newly developed for Regenova

Jul, 27, 2015 | Shipments of Regenova to North America set to start in September 2015

Jun, 30, 2015 | An article about Cyfuse on Bio Spectrum Asia, “Engineered 3D human tissue will bring efficiencies in drug screening and drug safety testing”

  1. Cyfuse Biomedical K.K. 3D bioprinter
  2. Bio 3D Printer Regenova, bioprinting