IP Library Granted Patent US 12,427,229
Granted Patent B2
US 12,427,229 · App. 17/387,801 · Granted Sep 30, 2025

Laser ablation/removal and laser induced forward transfer of biological material

Inventors: Ioanna Zergioti (Athens, GR); Apostolos Klinakis (Athens, GR)
Assignee: PhosPrint P.C.
A61L27/3691A61B18/20A61L27/28A61L27/3604A61L27/3679B41M3/006A61B2017/00969A61B2018/00494A61B2018/00505A61B2018/00577
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Quick Facts
Patent No.
US 12,427,229
App. No.
17/387,801
Granted
Sep 30, 2025
Kind
B2
Abstract

A method for cell printing is disclosed. The method includes generating a receiver substrate, ablating or removing a portion of the receiver substrate via a first laser to expose a target layer, generating a donor substrate containing a back surface and a front surface, applying a coating of donor material to the front surface. The method further includes aligning the front surface of the donor substrate to be parallel to and facing the receiver substrate, wherein the donor material is disposed adjacent to the target layer, and irradiating the coating through the back surface of the donor substrate with one or more laser pulses produced by a second laser to transfer a portion of the donor material to the target layer. A system for cell printing is also disclosed.

Claims (37)

1. A method comprising:

generating a receiver substrate, wherein the receiver substrate comprises intestinal tissue;

removing a portion of the receiver substrate via one or more lasers to expose a target layer of the receiver substrate for placement of donor material, wherein the donor material includes one or more cells suitable for proliferation on the target layer, wherein the donor material further comprises urothelial cells, wherein:

the urothelial cells comprise at least one of differentiated induced pluripotent stem cells (iPS) or stem cells of mesodermal or endodermal origin, or

the urothelial cells are derived from at least one of a bladder, a ureter, a urethra, or a renal pelvis;

generating a donor substrate, wherein the donor substrate comprises a back surface and a front surface;

applying a coating to the front surface, wherein the coating includes donor material;

aligning the front surface of the donor substrate to be parallel to and facing the receiver substrate, wherein the donor material is disposed adjacent to the target layer; and

irradiating the coating through the back surface of the donor substrate with one or more laser pulses produced by a second laser to transfer a portion of the donor material to the target layer, and

proliferating the one or more cells of the donor material on the target material to form a transplantable tissue.

2. The method of claim 1 , further comprising:

scanning the donor substrate through a focal point of the one or more laser pulses while irradiating the donor material with the one or more laser pulses to continuously provide new donor material to transfer to the receiver substrate; and

scanning the receiver substrate while irradiating the donor material with the one or more laser pulses to form a selected pattern of the donor material on the target layer.

3. The method of claim 2 , wherein the selected pattern of the donor material on the target layer comprises:

a layer of the donor material on the target layer.

4. The method of claim 1 , wherein removing the portion of the receiver substrate via the one or more lasers to expose the target layer of the receiver substrate comprises:

removing one or more crypt cells from the intestinal tissue without removing a stromal layer or a muscle layer from the intestinal tissue.

5. The method of claim 1 , wherein removing the portion of the receiver substrate via the one or more lasers to expose the target layer of the receiver substrate comprises:

denuding at least a portion of an epithelial layer of the intestinal tissue without removing a stromal layer or a muscle layer from the intestinal tissue.

6. The method of claim 1 , wherein the donor material comprises at least one of a tissue, a protein, a nucleic acid, an extracellular material, a scaffolding material, an epithelial cell, a urothelial cell, a fibroblast, a mesenchymal cell, an adipocyte, an immune cell, a muscle cell, a nerve cell, an insulinogenic cell, a keratinocyte, a chondrocyte or a stem cell.

7. The method of claim 1 , wherein the receiver substrate comprises at least one of an extracellular matrix, intestinal tissue, bladder tissue, stomach tissue, cartilaginous tissue, esophageal tissue, a cell-containing tissue, an organ, a portion of an organ, or an organoid.

8. The method of claim 1 , wherein the applying the coating comprises:

applying a dynamic release layer to the front surface of the donor substrate; and

applying the donor material to the dynamic release layer.

9. The method of claim 1 , wherein the removing a portion of the receiver substrate removes at least one of a layer of cells, an intestinal crypt, an extracellular matrix, a tissue, or portion of an organ from the receiver substrate.

10. A method comprising:

generating a receiver substrate, wherein the receiver substrate comprises biological tissue;

removing a portion of the receiver substrate via one or more lasers to expose a target layer of the receiver substrate for placement of donor material, wherein the donor material includes one or more cells suitable for proliferation on the target layer, wherein removing the portion of the receiver substrate via the one or more lasers to expose the target layer of the receiver substrate comprises:

removing one or more crypt cells from the intestinal tissue without removing a stromal layer or a muscle layer from the intestinal tissue or

or denuding at least a portion of an epithelial layer of the intestinal tissue without removing a stromal layer or a muscle layer from the intestinal tissue;

generating a donor substrate, wherein the donor substrate comprises a back surface and a front surface;

applying a coating to the front surface, wherein the coating includes donor material;

aligning the front surface of the donor substrate to be parallel to and facing the receiver substrate, wherein the donor material is disposed adjacent to the target layer; and

irradiating the coating through the back surface of the donor substrate with one or more laser pulses produced by a second laser to transfer a portion of the donor material to the target layer;

scanning the donor substrate through a focal point of the second laser while irradiating the donor material with the second laser to continuously provide new donor material to transfer to the receiver substrate; and

scanning the receiver substrate while irradiating the donor material with the second laser to form a selected pattern of the donor material on the target layer.

11. The method of claim 10 , wherein the receiver substrate comprises at least one of an extracellular matrix, intestinal tissue, bladder tissue, stomach tissue, cartilaginous tissue, esophageal tissue, a cell-containing tissue, an organ, a portion of an organ, or an organoid, wherein the donor material comprises at least one of a tissue, a protein, a nucleic acid, an extracellular material, a scaffolding material, an epithelial cell, a urothelial cell, a fibroblast, a mesenchymal cell, an adipocyte, an immune cell, a muscle cell, a nerve cell, an insulinogenic cell, a keratinocyte, a chondrocyte or a stem cell.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 30, 2021
From: ZERGIOTI, IOANNA; KLINAKIS, APOSTOLOS
To: PHOSPRINT P.C.
Reel/Frame 057031/0162 →
Continuity (2)
Provisional Application 63062176 · Aug 6, 2020
Related Publication 20220040377A1 · Feb 10, 2022
References Cited (59)
US 6905738B2 · Ringeisen et al. · 2005 [cited by applicant]
US 6936311B2 · Ringeisen et al. · 2005 [cited by applicant]
US 7001467B2 · Piquéet al. · 2006 [cited by applicant]
US 7507422B2 · Kropp et al. · 2009 [cited by applicant]
US 7875324B2 · Barron et al. · 2011 [cited by applicant]
US 9039998B2 · Guillemot · 2015 [cited by examiner]
US 9328327B2 · Haverich · 2016 [cited by applicant]
US 9629989B2 · Guillemot et al. · 2017 [cited by applicant]
US 9855369B2 · Murphy et al. · 2018 [cited by applicant]
US 10112388B2 · Guillemot · 2018 [cited by applicant]
US 10196596B2 · Glazier et al. · 2019 [cited by applicant]
US 20090246247A1 · Shetty et al. · 2009 [cited by applicant]
US 20110250688A1 · Hasan · 2011 [cited by applicant]
US 20110313429A1 · Anderson · 2011 [cited by examiner]
US 20150351896A1 · D'Lima et al. · 2015 [cited by applicant]
US 20160243286A1 · Collins et al. · 2016 [cited by applicant]
US 20160257926A1 · Rivron et al. · 2016 [cited by applicant]
US 20170196674A1 · Abdel-Meguid et al. · 2017 [cited by applicant]
US 20170320263A1 · Guillemot · 2017 [cited by applicant]
US 20170360551A1 · Liu · 2017 [cited by applicant]
US 20190328935A1 · Kang et al. · 2019 [cited by applicant]
US 20200055327A1 · Batt · 2020 [cited by applicant]
US 20200080060A1 · Matheu et al. · 2020 [cited by applicant]
US 20200102529A1 · Guillemot et al. · 2020 [cited by applicant]
AU 2002243924A1 · 2002 [cited by applicant]
CZ 1641955 · 2012 [cited by applicant]
EP 2542659B1 · 2018 [cited by applicant]
EP 3234102B1 · 2018 [cited by applicant]
EP 3526007A1 · 2019 [cited by applicant]
FR 3063930B1 · 2019 [cited by applicant]
RU 2323688C1 · 2008 [cited by applicant]
RU 2371102C1 · 2009 [cited by applicant]
RU 2408305C1 · 2011 [cited by applicant]
UA 53768C2 · 2003 [cited by applicant]
UA 55215A · 2003 [cited by applicant]
WO 2016124708A1 · 2016 [cited by applicant]
WO 2017187114A1 · 2017 [cited by applicant]
WO 2018193446A1 · 2018 [cited by applicant]
WO 2018193454A1 · 2018 [cited by applicant]
WO 2019060518A1 · 2019 [cited by applicant]
WO 2019122351A1 · 2019 [cited by applicant]
WO WO2019129349A1 · 2019 [cited by examiner]
WO 2019198086A1 · 2019 [cited by applicant]
Serrano-Aroca et al. Bioengineering Approaches for Bladder Regeneration. International Journal of Molecular Sciences. Vo. 19. 1796. 2018 (Year: 2018). [cited by examiner]
Fraser, M. et al., (2004), “A surgical model of composite cystoplasty with cultured urothelial cells: a controlled study of gross outcome and urothelial phenotype”, BJU International, 93: 609-616. doi:10.1111/j.1464-410… [cited by applicant]
Jonason et al., Manuscript, “Primary Murine Growth Plate and Articular Chondrocyte Isolation and Cell Culture” published in Osteoporosis and Osteoarthritis on Sep. 17, 2014, 8 pages. [cited by applicant]
Kalabis et al., Manuscript, “Isolation and characterization of mouse and human esophageal epithelial cells in 3D organotypic culture” published in Nature Protocols, on Jan. 12, 2012, 12 pages. [cited by applicant]
Munaz, A. et al., “Three-dimensional printing of biological matters”, Journal of Science: Advanced Materials and Devices, 1 (2016) 1-17. [cited by applicant]
Murphy, S. et al., “3D bioprinting of tissues and organs”, Nat Biotechnol 32, 773-785 (2014). https://doi.org/10.1038/nbt.2958. [cited by applicant]
Nik et al., Manuscript, “Separation of Intact Intestinal Epithelium from Mesenchyme” published in Biotechniques on Jul. 2014, 3 pages. [cited by applicant]
Oberpenning, F. et al., “De novo reconstitution of a functional mammalian urinary bladder by tissue engineering”, Nat Biotechnol 17, Abstract Only (1999). https://doi.org/10.1038/6146. [cited by applicant]
Precise Bio, URL: https://www.precise-bio.com/, downloaded Apr. 24, 2020, 3 pages. [cited by applicant]
Prellis Biologics, URL: https://www.prellisbio.com/tech, downloaded Apr. 24, 2020, 1 page. [cited by applicant]
Oberpenning, F. et al., “De novo reconstitution of a functional mammalian urinary bladder by tissue engineering”, Nat Biotechnol 17, (1999). https://doi.org/10.1038/6146. [cited by applicant]
European Patent Office, International Search Report and Written Opinion for International Application No. PCT/GR2021/000052, Nov. 26, 2021, 18 pages. [cited by applicant]
Higbee et al., “Femtosecond Laser Ablation of Porcine Intestinal Mucosa: Potential Autologous Transplant for Segmental Cystectomy,” Proceedings of SPIE, vol. 5686, Photonic Therapeutics and Diagnostics, Apr. 25, 2005, 1… [cited by applicant]
Koo et al., “Laser-assisted Biofabrication in Tissue Engineering and Regenerative Medicine,” JMR Early Career Scholars in Materials Science Annual Issue: Review, vol. 32, No. 1, Dec. 19, 2016, 15 pages. [cited by applicant]
Pere Serra et al., “Laser-Induced Forward Transfer: Fundamentals and Applications,” Advanced Materials Technologies, vol. 4, No. 1, Aug. 8, 2018, 33 pages. [cited by applicant]
Serrano-Aroca et al., “Bioengineering Approaches for Bladder Regeneration,” International Journal of Molecular Sciences, vol. 19, No. 6, Jun. 17, 2018, 26 pages. [cited by applicant]