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‘Cryobioprinting’ serves up towers of frozen

Photo: Cryobioprinting Examples
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Credit: Credit Y. Shrike Zhang

A new technology takes bioprinting – in which cell ink is printed, layer by layer, to form a structure – to a whole new, icy level. Investigators from Chang’s lab at Brigham and Women’s Hospital have developed a technique they call “cryobioprinting,” a method that uses a bio-linker embedded with cells to print frozen, complex structures that can easily be stored for later use. The team presented the cryobioprinting technique in a paper recently published in Theme It also described how the technology is applied to muscle tissue engineering in a research paper just published in Sophisticated materials.

“Cryolipolysis can give bioprinted tissues a long shelf life. We’ve shown up to three months of storage, but it can be much longer,” said Y. Shrike Zhang, Ph.D., lead author on both papers and an associate bioengineer in Brigham’s Department of Medicine. “And the unique variation, or what we call the vertical 3D cryolithography technology that we describe, may have broad applications in tissue engineering, regenerative medicine, drug discovery, and personalized therapies.”

Zhang and colleagues used a cryo-protected biolinker loaded with cells to print tissue constructs on a custom freezing plate. The freeze plate allowed them to precisely control and hold the temperature while the cryogenic printing was being done. These printed structures were immediately kept in a liquid nitrogen tank for later use. The team refined and evaluated the technology, finding that it could faithfully manufacture tissue formulations that could be used as implants and tissue products.

in a advanced materialsZhang and co-authors report the use of the cryoprotected biolinker to create 3D vertical structures that mimic the complex, delicate, and anisotropic tissues found in the human body. Many tissues of the body, including muscles and nerve cells, are anisotropic, which means that they have different properties in different directions. The structures created by the researchers were also anisotropic, with micropores aligned in the vertical direction. As a proof of concept, the team built a muscle tendon unit using myocytes (cells that can give rise to muscle cells) and fibroblasts (cells that produce the skeletal frameworks in connective tissue). The team also made the muscle’s microvascular unit.

The researchers note that this work represents very early technology demonstrations and will still need validation and extensive testing before it can be used in the clinic, but the two papers represent an important step forward.

“As the field of tissue engineering is rapidly growing, these manufactured tissue formulations may find a large number of applications in muscle tissue engineering and beyond,” Zhang said.

Financing: The authors acknowledge the support of the Brigham Research Institute. The work was also supported by FRQNT’s International Internship Award (279390), MITACS Globalink Research Award (IT14553), McGill’s Graduate Mobility Award, McGill’s Doctor Internship Award, FRQNT’s Postdoctor Fellowship (296447), Program of China Scholarship Award (No.201807045057), Henan Provincial High Level Talent Internationalization Training Program (No. 20199004), National Institute of Deafness and Other Communication Disorders (NIDCD) of the National Institutes of Health (NIH) Grant Numbers R01DC005788 and R01DC014461.

Papers cited:

Ravanbakhsh H et al. Free-form cell-loaded cryoprinting for the manufacture and storage of finished tissues. Theme DOI: /10.1016/j.matt.2021.11.020

Luo Z et al. Support for bath-free vertical (bio) extrusion cryoprinting for the fabrication of anisotropic tissues. advanced materials DOI: 10.1002 / adma.202108931

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