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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Results
  • Discussion
  • Disclosures
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

Soluble basement membrane extracts are the most widely used biological matrices in cancer research, but their complex rheological behaviour makes their bioprinting difficult with commercially available bioprinting systems. This work presents a customized bioprinting strategy to produce pure matrix constructs with good shape fidelity in both single and multiple layers.

Abstract

In translational research, 3D in vitro models are essential for advancing scientific understanding and therapeutic development. To this aim, Soluble Basement Membrane Extracts (SBMes), derived from mouse sarcoma cells, play the key role of replicating mechanical and biochemical features of in vivo microenvironment. They are particularly valuable in a variety of research fields, from tissue engineering to drug testing. In particular, they have become fundamental in cancer research as traditional 2D cell cultures, widely used in the field, can provide misleading information since they do not capture the three-dimensional (3D) structure and tumor microenvironment.

In cancer research, effective 3D in vitro models are crucial for better understanding cancer evolution and anticipating challenges, such as drug resistance mechanisms. Organoids have emerged as promising 3D in vitro models, offering a more accurate representation of tumor biology. They grow in SBMes as they create an environment where cells can grow and self-organize into 3D structures. However, SBMe presents significant challenges, including low mechanical properties and complex rheological behavior, preventing its use with commercially available pneumatic-extrusion bioprinting systems.

To address these limitations, we developed a low-cost, volumetric-controlled bioprinting system and a specific protocol for printing structures with SBMe. This system is based on a volumetric extruder and overcomes the limitations of traditional pneumatic-driven approaches. Once assembled and programmed with a specific g-code, the system can bioprint both pure and diluted SBMe to obtain both single and multi-layer constructs. This approach offers a more reliable and scalable method for producing 3D cell culture models, paving the way for more effective research into novel treatments and drug testing.

Introduction

In translational research, creating 3D in vitro models that closely replicate the in vivo environment is essential for advancing scientific understanding and therapeutic development1. A key component in building them is the use of extracellular matrix (ECM) analogs, such as Soluble Basement Membrane Extracts (SBMes), which better reflect the complexities of natural tissues. SBMes are particularly valuable in a variety of research fields, from tissue engineering to drug testing2,3. In particular, they have become fundamental in cancer research, as they replicate mechanical and biochemica....

Protocol

1. 3D printing and assembly of volumetric bioprinter

  1. Prepare and 3D-print the bioprinter components.
    1. Download the *.STL design files from Supplemental File 1.
    2. Load *.STL file in the proprietary slicing software to define the orientation of the pieces on the printing plate and all the options required for the 3D printer. Print the pieces using polylactic acid (PLA) with medium-quality options (0.4 mm nozzle size, 1.75 mm filament diameter, 220 Β°C extruder temperature, 50 Β°C print bed plate temperature, 0.2 mm layer height, 300 mm/s printing speed).
      NOTE: Medium-quali....

Results

In this article, we present a protocol for bioprinting constructs made of SBMe using a low-cost, custom-made, volumetric bioprinter. We provide a detailed description and 3D printing *.STL files to build the system and we demonstrate its potential use in cancer research by printing single and multi-layer structures. Each 3D printed component has been properly cleaned from support structure or unwanted printing residues before assembling. Figure 8 shows different views of the overall assembly.......

Discussion

In translational research, SBMes are particularly valuable in a variety of research fields, from tissue engineering to drug testing2,3. In particular, they have become fundamental in cancer research, as they replicate mechanical and biochemical features of the tumor microenvironment, enabling the development of models that better reflect tumor growth, metastasis, and treatment responses. SBMe is the most widely used biological matrix, especially for 3D organoids<.......

Disclosures

The authors have no conflicts of interest to declare.

Acknowledgements

This work was partially funded by the Accelerator Award nΒ° A26815 entitled: "Single-cell cancer evolution in the clinic" funded through a partnership between Cancer Research UK and Fondazione AIRC (nΒ° 22790) and by the European Union - Next Generation EU, Mission 4, Component 1, CUP D53D23003310006.

....

Materials

NameCompanyCatalog NumberComments
12-multiwell plateSigma-AldrichCLS351143
3D FDM printerFlashforgeAdventurer 5M Pro
3D printer MotherboardGEETECHGT2560 REV A+
6-multiwell plateSigma-AldrichM8687
AC socketHiLetgohttps://www.amazon.it/HiLetgo-Terminal-Socket
-Holder-Switch/dp/B0814PT3YG/ref=sr_1_1?__
mk_it_IT=%C3%85M%C3%85%C5%BD%C3%
95%C3%91&crid=2TRIBP34VJ61A
&dib=eyJ2IjoiMSJ9.zW6_VmeiudR
I4yMpjJqJm7OdeoctTKW2mrEzml
BAyJVa1hkM4PiJT3pLY0mXdxm1
AZQxa2_f5J4q4d5v3vUeqLkjJ6BM
kWHAoeEfbEQJNuZaQ3NjmUlqWy
_1AfQpnRp4VqJ2m3bkWsChztCa
Ok-ZdQJlobGxJaMKg7WbV352
Hyo.0tEw2GYoPQJ4SdNYXO1XM
Jrl2yXMvRey55Hj7GhISkk&dib_
tag=se&keywords=AC+socket+HiLetgo&nsdOpt
OutParam=true&qid=1734702097&sprefix=ac+
socket+hiletgo%2Caps%2C173&sr=8-1		15 A 250 V
Adjustable feetEXIN DEHCENhttps://www.amazon.it/gp/product/
B0CPLTSNQB/ref=ppx_yo_dt_b_
search_asin_title?ie=UTF8&psc=1		Height 27-40 mm
Ball bearing 4 mmQUARKZMAN684ZZDimensions 4 x 9 x 4 mm
Ball bearing 5 mmXiKe625-2RSDimensions 5 x 16 x 5 mm
Belt closedTurmberg3Dhttps://www.amazon.it/gp/product/
B09B2FJQBV/ref=ppx_yo_dt_b_
search_asin_title?ie=UTF8&psc=1		Width 6 mm
Belt openFajoedahttps://www.amazon.it/gp/product/
B09QCTVPTH/ref=ppx_yo_dt_b_
search_asin_title?ie=UTF8&th=1		Width 6 mm
Bioprinting nozzleFisher Scientific17780789CorningΒ Standard Conical Bioprinting Nozzles
BΓΌrker Cell Counting chamberGLASWARENFABRIK KARL HECHT40443001
CentrifugeEppendorf5702 R
Centrifuge tubeSigma-AldrichCLS43147015 mL
Centrifuge tubeSigma-AldrichEP00301200861.5 mL
DisplaySigma-AldrichBR718905
DMEMΒ Gibco Thermofisher11965092
FBSSigma-AldrichF7524
Flanged linear ball bearingSourcing maphttps://www.amazon.it/gp/product/
B07S6LZHM7/ref=ppx_yo_dt_b_
search_asin_title?ie=UTF8&psc=1		LMF8UU
Glass syringeFisher Scientific11520062Hamilton GasTight 1 mL (TLL terminal)
HingesYASQZhttps://www.amazon.it/gp/product/
B09DNXGDGG/ref=ppx_yo_dt_b_
search_asin_title?ie=UTF8&psc=1
IncubatorFisher Scientific15481374EppendorfΒ Galaxy 48 R CO2Β Incubator, 48 L, Stainless Steel
Inverted microscopeNikonEclipse Ti2-U
Jumper wiresIverntechhttps://www.amazon.it/Iverntech-XH2
-54-Terminale-Stampante-Stepper/dp/
B07Q12B6K5/ref=pd_ci_mcx_mh_
mcx_views_0_image?pd_rd_w=cb2r
J&content-id=amzn1.sym.57a351cf-
ba07-47e8-b302-aa2aa1a3f209%3
Aamzn1.symc.ca948091-a64d-450e-
86d7-c161ca33337b&pf_rd_p=57a3
51cf-ba07-47e8-b302-aa2aa1a3f209
&pf_rd_r=YHY9RFYZJK7PYAY2RXX
D&pd_rd_wg=7WkXB&pd_rd_r=8239
66cb-4977-48ce-b095-0dcedf8c372f&
pd_rd_i=B07Q12B6K5&th=1
LCD unitParadisetronic.comhttps://www.amazon.it/Display-controller
-conduttori-adattatore-stampante/dp/B01
DUR4064/ref=sr_1_9?__mk_it_IT=%C3
%85M%C3%85%C5%BD%C3%95%C3
%91&crid=1OAHVIIRA16X3&dib=eyJ2Ij
oiMSJ9.07V2Jf7RzJjTUlIuFobUJFjUzG4
8633KkdcMXSEfnWLRX41d5vAF7xPBJ
maniVxDoXecfihZzzOXlP3v-e29cUjoGLf
GDyv5DoDVwp7ndohNTYvYzoi3gWkF-
Hsd2YbT7tjb1Dra1afqSn26CbFdEvJ84y
qyX7Gi3-7DHh8yMwdd6_7quiuW_mjsm
w1nu0YQG68GZ0XM7unOsvyddQ__R_
3dndxb3mOjgGjklEgu_KEbZJPOZDIdSo
oU3nrrKHj56hCfk9ACwYpB80QGCbTPK
0p-fE0g9h02WPmtHV38UU3m5AdZsTQ
HqvnVlJg703qGT-8ze8fQItw2rtbgl_J86H
RMReJOhVLQBkqNwxh57CQ.yNbeFmZ
ZBEUhadz2dQa8NxpZD9g4P6ighRU_97
yxgnw&dib_tag=se&keywords=display%
2B3D%2Bprinter%2Bprusa%2Bi3&nsdO
ptOutParam=true&qid=1734714093&s=
industrial&sprefix=display%2B3d%2B
printer%2Bprusa%2Bi3%2Cindustrial%
2C143&sr=1-9&th=1		Old Prusa i3 LCD unit. Link to commercially available alternative
Leadscrew M8RS PRO280-408
Leadscrew nut M8Comiokehttps://www.amazon.it/dp/B0C7QB13C4?
ref=ppx_yo2ov_dt_b_fed_asin_title&th=1		Nut for M8 screws
Leadscrew nut T8Aipaidehttps://www.amazon.it/gp/product/B086QJCX1M/ref
=ppx_yo_dt_b_search_asin_title?ie=UTF8&psc=1		Nut for T8 screws
Leadscrew T8VBESTLIFEhttps://www.amazon.it/gp/product/B07CXRB52Y/ref
=ppx_yo_dt_b_search_asin_title?ie=UTF8&th=1		Length 15 mm
L-GlutammineSigma-AldrichG7513
Limit switchCESFONJERhttps://www.amazon.it/gp/product/B07SPX492J/ref
=ppx_yo_dt_b_search_asin_title?ie=UTF8&psc=1
Linear ball bearing (LM4UU)A ABSOPROhttps://www.amazon.it/gp/product/B0CB3M5GHJ/ref
=ppx_yo_dt_b_search_asin_title?ie=UTF8&psc=1		Dimensions 4 x 8 x 12 mm
Linear ball bearing (LM8LUU)Fdithttps://www.amazon.it/LM8LUU-Linear
-Motion-cuscinetti-stampante/dp/B07N
58W3GZ/ref=sr_1_1_sspa?__mk_it_IT
=%C3%85M%C3%85%C5%BD%C3%
95%C3%91&crid=33VNCCCSIFJRP&
dib=eyJ2IjoiMSJ9._JJ2doXI33C2JagX
8x6E2Bz2ILo10852tBi1erWqGn363ku
zbcvknbMtsK6wm5jdR1P5XwC6Ovui
at3kGIJPpYFOG_FiIBYT8xLkkbiphhR
4ncR6R9tPPyeN--_3RxFgKMh5tMO_
jesAcu7Anp87Qb91ruM85CKqduV-y6
HQJtAvVSXmfG4yE4N7d4_dDVKsR7
UkEmDIznJE0HIhvvI2R6vX2tMN5yMc
_ZrUbUB0gPF6SgXHVJmKRizCRqL7J
64WRD-XnMSpVHY_LUVJ7gn9kJQ35
_zQw8vLD-_kRyvxwvE.4IVMdKzF1Ax
mQHQhWEi6LH12UI4CmMgi288_g3zG
CPY&dib_tag=se&keywords=LM8LUU+
Fdit&nsdOptOutParam=true&qid=17347
04494&s=industrial&sprefix=lm8luu+fdit+
%2Cindustrial%2C136&sr=1-1-spons&sp
_csd=d2lkZ2V0TmFtZT1zcF9hdGY&psc=1		Dimensions 8 x 15 x 45 mm
Linear ball bearing (LM8UU)ARCELIhttps://www.amazon.it/gp/product/
B07BV3YBP2/ref=ppx_yo_dt_b_
search_asin_title?ie=UTF8&psc=1		Dimensions 8 x 15 x 24 mm
MagnetsOCEUMAOAhttps://www.amazon.it/gp/product/
B0CB6C2RF4/ref=ppx_yo_dt_b_
search_asin_title?ie=UTF8&th=1		8 x 3 mm
MatrigelGibco ThermofisherLOT 0202003SBMe
Motor driverΒ Longrunerhttps://www.amazon.it/gp/product/
B071P41ZBW/ref=ppx_yo_dt_b_search
_asin_title?ie=UTF8&psc=1		Driver A4988
PBSMicroGem, VWRTL1006-500ML
Penicillin-StreptomycinGibco Thermofisher15070063
Pipette tipsSigma-AldrichZ740058BRANDΒ filter tips, racked, TipBox (volume 50-1000 ΞΌL)
Plexiglass sheetsMVQPERhttps://www.amazon.it/MVQPER-trasparenti-
plexiglass-trasparente-decorazione/dp/B0CF
Q4BHQZ/ref=sr_1_14?dib=eyJ2IjoiMSJ9.bf7
Mp7I5m2qAR1Fp1V6DoYF2erUlNQUK_NuF
ZLz9A7uJKGa_RNeni6NiO4YQfAjVHBtZgdzI
356XVc3BIuKVpV8Pfg0rImqNXgxG-kfklAYrb
rXv4tV6RL1ShFwgLzfQh8R0sEoFJPcLF2f2E
EcGMlvEwOuCv6qRnS1MQ-VaA0sLPnM0ts
wdL4sLVxsonWcfSrHAUrgWh4fpVNMA4Kwh
0UaeQhfexFY0dNWhPYftXvvz9LLj1uMID2E
Gi_ScmYqY5LahbYOmKVVPPiPIgCVdLjaM8
yqNp2ZJGV9Fge2xa6OAHxmMeaQUqGkoQ
88x9GkX7EWuOwYO0dEUYSzHCdGONWo
eZLfSJOR3C6-knjzKrOQ0ziouJfo-ktbak7v3A
H53BtIOiXbT57cKqh-uc9wI_j_lHN-8okAe2yO
6upc61MoydZujuxnfLG7hwytqHQG2.ApKq8
MVs8Xihn_zcA39nML-2lpjnagGhXIx0clcJN_
E&dib_tag=se&keywords=lastra+plexiglass+
2+mm&nsdOptOutParam=true&qid=1734702823&sr=8-14		2 mm thickness, transparent sheet, 21 x 32.5 cm
Plexiglass sheetsMVQPERhttps://www.amazon.it/MVQPER-trasparenti
-plexiglass-trasparente-decorazione/dp/B0C
FQ4BHQZ/ref=sr_1_14?dib=eyJ2IjoiMSJ9.b
f7Mp7I5m2qAR1Fp1V6DoYF2erUlNQUK_N
uFZLz9A7uJKGa_RNeni6NiO4YQfAjVHBtZ
gdzI356XVc3BIuKVpV8Pfg0rImqNXgxG-kfk
lAYrbrXv4tV6RL1ShFwgLzfQh8R0sEoFJPc
LF2f2EEcGMlvEwOuCv6qRnS1MQ-VaA0s
LPnM0tswdL4sLVxsonWcfSrHAUrgWh4fpV
NMA4Kwh0UaeQhfexFY0dNWhPYftXvvz9
LLj1uMID2EGi_ScmYqY5LahbYOmKVVPP
iPIgCVdLjaM8yqNp2ZJGV9Fge2xa6OAHx
mMeaQUqGkoQ88x9GkX7EWuOwYO0dE
UYSzHCdGONWoeZLfSJOR3C6-knjzKrO
Q0ziouJfo-ktbak7v3AH53BtIOiXbT57cKqh-
uc9wI_j_lHN-8okAe2yO6upc61MoydZujux
nfLG7hwytqHQG2.ApKq8MVs8Xihn_zcA3
9nML-2lpjnagGhXIx0clcJN_E&dib_tag=se
&keywords=lastra+plexiglass+2+mm&nsd
OptOutParam=true&qid=1734702823&sr=8-15		2 transparent sheets, 2 mm thickness, 20 x 20 cm
Plexiglass sheetsMVQPERhttps://www.amazon.it/MVQPER-trasparenti
-plexiglass-trasparente-decorazione/dp/B0C
FQ4BHQZ/ref=sr_1_14?dib=eyJ2IjoiMSJ9.
bf7Mp7I5m2qAR1Fp1V6DoYF2erUlNQUK_
NuFZLz9A7uJKGa_RNeni6NiO4YQfAjVHB
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0sLPnM0tswdL4sLVxsonWcfSrHAUrgWh
4fpVNMA4Kwh0UaeQhfexFY0dNWhPYft
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2xa6OAHxmMeaQUqGkoQ88x9GkX7E
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N_E&dib_tag=se&keywords=lastra+plexigl
ass+2+mm&nsdOptOutParam=true&qid=1
734702823&sr=8-16		2 transparent sheets, 2 mm thickness, 20 x 30 cm
Polylactic acid (PLA) filamentFlashforgeBlack 100102
Power supplySQUADOhttps://www.amazon.it/SQUADO-Alimentatore
-stabilizzato-dissipatore-efficienza
/dp/B0DMTG6SJD/ref=sr_1_6?dib
=eyJ2IjoiMSJ9.1vVze6Zt6fhKCvFb
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C-iqBEu-vjIVgsrQmOjeLuGo9YQ&
dib_tag=se&keywords=power+sup
ply+AC+INPUT%3A+110-220V%2
C+DC+OUTPUT%3A+12V+15A&n
sdOptOutParam=true&qid=173471
4722&sr=8-6		Old Prusa i3 power supply (AC INPUT: 110-220 V, DC OUTPUT: 12 V 15 A). Link to commercially available alternative
Pulley 5 mmVooGenzekhttps://www.amazon.it/gp/product/
B0B8H16KWX/ref=ppx_yo_dt_b_
search_asin_title?ie=UTF8&psc=1		Bore 5 mm, Width belt space 6 mm
Pulley 8 mmYxtaiihttps://www.amazon.it/gp/product/
B07X852RFN/ref=ppx_yo_dt_b_
search_asin_title?ie=UTF8&th=1		Bore 8 mm, Width belt space 6 mm
Ribbon cablesQUARKZMANhttps://www.amazon.it/sspa/click?ie
=UTF8&spc=MTo0NDgyNTkwNzU
wNjM5NzM5OjE3MzQ3MTQ1OTE6
c3BfbXRmOjMwMDEyMjk4NTMzN
TYzMjo6MDo6&url=%2FQUARKZM
AN-connettore-Lunghezza-Comput
er-confezione%2Fdp%2FB0CQRM
DQNG%2Fref%3Dsr_1_15_sspa%
3F__mk_it_IT%3D%25C3%2585M
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C3%2595%25C3%2591%26crid%3
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sdOptOutParam%3Dtrue%26qid%
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%26sprefix%3Dcavo%2Bflessibile
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%26sr%3D1-15-spons%26sp_csd%
3Dd2lkZ2V0TmFtZT1zcF9tdGY%26psc%3D1		ribbon cable 2651 28AWG 10 pins
Rigid shaft couplingGwolfhttps://www.amazon.it/gp/product/
B088R8CW5Z/ref=ppx_yo_dt_b_
search_asin_title?ie=UTF8&psc=1		Coupling from 5 to 8 mmΒ 
Rods 4 mmRS-components682-810Diameter 4 mm
Rods 5 mmSourcingmapa14010200ux0214Diameter 5 mm
Rods 8 mmRS-components682-078Diameter 8 mm
ScrewsBnnrjia281-984M2x6 - Flat head
ScrewsINCREWAYhttps://www.amazon.it/INCREWAY-
assortite-portatili-accessori-
riparazione/dp/B07KN37M5J/
ref=sr_1_20?__mk_it_IT=%C
3%85M%C3%85%C5%BD%
C3%95%C3%91&crid=12BH
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0R.jCyNdxQiJSAsGVm7ZPhDK
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ib_tag=se&keywords=viti+M3x3&
nsdOptOutParam=true&qid=1734
711519&sprefix=viti+m3x3%2Cap
s%2C169&sr=8-20		M2x8 - Flat head
ScrewsINCREWAYhttps://www.amazon.it/INCREWAY
-assortite-portatili-accessori-ripara
zione/dp/B07KN37M5J/ref=sr_1_2
0?__mk_it_IT=%C3%85M%C3%85
%C5%BD%C3%95%C3%91&crid=
12BHMAWD8SK4Z&dib=eyJ2IjoiM
SJ9.SjXKoxV0NYqENPoUeS42-rj1
Af398uiCLXDxFs0C9HfS8vwEPms
fAMPwIMfllkAfWNYTIEU4KXhJRg
RHInNI3U_aow0EPyfufQKlA4wHz
1eOJBvYOND-5obbXfRPn7VXQ8
kzBXMVfUHLAIFlJhTO8AXclkwgjB
-zpRvQ6oE5Fr_1sO7lFnzURtD1YB
f65O58M3hrDXf5r_8MIz1cAsct-2P8
AyWWBZAB1buYoyPgnnOiBL5EBS
7twJWqcDU86mh1ywyufiUitm6kIwN
uMx32D0h0jkHna481aaYP_Q7kHg
DxxrR_fFXj5gV2XqEesqGkxPCmHf
FWVMqvB_YBdEAuTeL08EP_pTY50
uT4UeWgdJuT58kB7Gv-Y5fPMPRS
_JEQdO-oJ9DlO9n0OszkmYwrPh-
AivwVN6xh4hN2pseXannS_on9aiN
v1lh3x-f2PZ0R.jCyNdxQiJSAsGVm
7ZPhDKeFxQGBAu-M_42ZLTSSB
iDg&dib_tag=se&keywords=viti+M3
x3&nsdOptOutParam=true&qid=17
34711519&sprefix=viti+m3x3%2Ca
ps%2C169&sr=8-20		M2x10 - Flat head
ScrewsINCREWAYhttps://www.amazon.it/INCREWAY-
assortite-portatili-accessori-riparazi
one/dp/B07KN37M5J/ref=sr_1_20?
__mk_it_IT=%C3%85M%C3%85%
C5%BD%C3%95%C3%91&crid=12
BHMAWD8SK4Z&dib=eyJ2IjoiMSJ
9.SjXKoxV0NYqENPoUeS42-rj1Af3
98uiCLXDxFs0C9HfS8vwEPmsfAM
PwIMfllkAfWNYTIEU4KXhJRgRHIn
NI3U_aow0EPyfufQKlA4wHz1eOJB
vYOND-5obbXfRPn7VXQ8kzBXMV
fUHLAIFlJhTO8AXclkwgjB-zpRvQ6
oE5Fr_1sO7lFnzURtD1YBf65O58
M3hrDXf5r_8MIz1cAsct-2P8AyWW
BZAB1buYoyPgnnOiBL5EBS7twJ
WqcDU86mh1ywyufiUitm6kIwNuM
x32D0h0jkHna481aaYP_Q7kHgDx
xrR_fFXj5gV2XqEesqGkxPCmHfF
WVMqvB_YBdEAuTeL08EP_pTY5
0uT4UeWgdJuT58kB7Gv-Y5fPMPRS
_JEQdO-oJ9DlO9n0OszkmYwrPh-Ai
vwVN6xh4hN2pseXannS_on9aiNv1l
h3x-f2PZ0R.jCyNdxQiJSAsGVm7Z
PhDKeFxQGBAu-M_42ZLTSSBiDg
&dib_tag=se&keywords=viti+M3x3&n
sdOptOutParam=true&qid=1734711
519&sprefix=viti+m3x3%2Caps%2
C169&sr=8-20		M3x3 - Flat head
ScrewsRS-components281-978M3x6 - Flat head
ScrewsRS-components281-978M3x10 - Flat head
ScrewsRS-components281-978M3x20 - Flat head
ScrewsFaankitonhttps://www.amazon.it/Faankiton-600-
M3-Incassato-Cilindrica/dp/B0CFY6V
3HV/ref=sr_1_3_sspa?crid=29NMQZ
B1QM7EW&dib=eyJ2IjoiMSJ9.aiGS-
zo819G6hZmghT7TFDH1TL5lzRsjT
R5PBvqoHbKOBLipyX-kuevmqqPpW
m_7Ngng-Wa6aLoDuXMtoqexgeNpr
Qn2cISsCatqzhc8fxhJ0yIN4s3YEpD
KzIroWWVNzswptkWthnGkLXwHck
m4wqLr1sf0qyK9oYYlZdKo_Ypa9RQ
s3aYPNd_f-DWpT5rdsG1GL3rd3b9c
fLSqXxMd8QnnK3hDcg10Uz9otPwo
zJC2QAQFce__CkNwDfBp5Vvgmy1
DLX4a7PolnNePrJiObgjHMJLlfwizGx
fie28RAYQ.03Epc_RrVWlBUqUbHU
TPDcoDUxaKBHep1CuTOhFj-fI&dib
_tag=se&keywords=m3x25%2Bviti&n
sdOptOutParam=true&qid=17347118
59&s=industrial&sprefix=M3x25%2Ci
ndustrial%2C185&sr=1-3-spons&sp_
csd=d2lkZ2V0TmFtZT1zcF9hdGY&th=1		M3x25 - Flat head
ScrewsFaankitonhttps://www.amazon.it/Faankiton-600
-M3-Incassato-Cilindrica/dp/B0CFY6
V3HV/ref=sr_1_3_sspa?crid=29NMQ
ZB1QM7EW&dib=eyJ2IjoiMSJ9.aiG
S-zo819G6hZmghT7TFDH1TL5lzRsj
TR5PBvqoHbKOBLipyX-kuevmqqPp
Wm_7Ngng-Wa6aLoDuXMtoqexgeN
prQn2cISsCatqzhc8fxhJ0yIN4s3YEp
DKzIroWWVNzswptkWthnGkLXwHc
km4wqLr1sf0qyK9oYYlZdKo_Ypa9R
Qs3aYPNd_f-DWpT5rdsG1GL3rd3b9
cfLSqXxMd8QnnK3hDcg10Uz9otPwo
zJC2QAQFce__CkNwDfBp5Vvgmy1D
LX4a7PolnNePrJiObgjHMJLlfwizGxfie
28RAYQ.03Epc_RrVWlBUqUbHUTPD
coDUxaKBHep1CuTOhFj-fI&dib_tag=s
e&keywords=m3x25%2Bviti&nsdOptO
utParam=true&qid=1734711859&s=ind
ustrial&sprefix=M3x25%2Cindustrial%2
C185&sr=1-3-spons&sp_csd=d2lkZ2V
0TmFtZT1zcF9hdGY&th=1		M3x30 - Flat head
ScrewsAERZETIXhttps://www.amazon.it/AERZETIX-
carbonio-fissaggio-cacciavite-mont
aggio/dp/B0BDYZZ8LQ/ref=sr_1_4
?__mk_it_IT=%C3%85M%C3%85
%C5%BD%C3%95%C3%91&crid=
217010XNB7ZXP&dib=eyJ2IjoiMSJ
9.e5QjA-q9hEvlleIzwY3ssA3x7vIBF
Hpcg73tKkBJu9QbDwMwlC5L9-9u
7my7ZG5hZij0WyaijakqAoX4rIZ3rv
EgvlPtN6bwKTBgqAK-3uu3GBy6K
EPQ2iIEUWx_U73nonJy5IzmTDH
Xijn-kRg1iCOM99S5upSWbmmazg
8lctsyXlg_tmXHjHn63WsIo7WcBuL
rtVpPW8oMsZCndwh33pNQXBnk1
iStT81spopzrw3gScK1Wdz6RttH6j
YL611rNBdkd-PEP2CYhDKeSKHE
AqmABjHGLyG6p40k20npHqkg8g-
9Msl3fZZiS4rRxNFOHBjEdVTLBrn
PjRjxTspBfZSEKQnqsGm1WPlz8cS
khCs.ujWHAL4y-RNahay-1AJl34uN
hUj-f6mZ7X1_KZaACvU&dib_tag=s
e&keywords=m3x50%2Bviti&nsdOp
tOutParam=true&qid=1734711964&
s=industrial&sprefix=m3x50%2Bviti
%2Cindustrial%2C136&sr=1-4&th=1		M3x50 - Flat head
ScrewsJOCAHULFXhttps://www.amazon.it/Incassato-
Esagonale-Rondelle-Esagonali-
Inossidabile/dp/B0DF7P5CC2/ref=sr_
1_6?__mk_it_IT=%C3%85M%C3%8
5%C5%BD%C3%95%C3%91&crid=2
7B231M7SRJ3R&dib=eyJ2IjoiMSJ9.3
bij8nHLQAebjY8LHQS6V1GsAE07H
cFGUpHAh7jA1oQIxjoTJmxjNuwg0y
H14oz-YUvzD0eqVRASqAZTK2spiQ
saX0pug7rWgGfENeGhztno5lMcJDT
XQfe9H6ocLZANpAFv_p0dRbO6ggA
vZN0TfyiuR4focFMAjr6VFGVbprEQO
EaS1KRs8loF3FgL0rsmP81K806Gv5
zeSurK_sp8GEelqtm1G3J059zUdPaJ
z7zJQEFPSH0TXv3lml5u9lnvnAbM0j
ClF03nG5ZjB-6WUPfS41Os5DgzTpS
zUTR-YZc1HnRUxy6l5grDPFmDiNZn
0Ie9MrlkcPI2A8dubQZm3d8pYVpg6Y
2OqzifmZzNsQM.Zuh5Ko4bf20Bz46c
NFRHJvcyPfJyglpcvN1uM16-yOI&dib
_tag=se&keywords=m4+viti+testa+sv
asata&nsdOptOutParam=true&qid=17
34712054&s=industrial&sprefix=m4+v
iti+testa+svasata%2Cindustrial%2C12
8&sr=1-6		M4x8 - Flat head
ScrewsJOCAHULFXhttps://www.amazon.it/Incassato-Esagonale
-Rondelle-Esagonali-Inossidabile/dp/B0DF
7P5CC2/ref=sr_1_6?__mk_it_IT=%C3%85
M%C3%85%C5%BD%C3%95%C3%91&c
rid=27B231M7SRJ3R&dib=eyJ2IjoiMSJ9.
3bij8nHLQAebjY8LHQS6V1GsAE07HcFG
UpHAh7jA1oQIxjoTJmxjNuwg0yH14oz-YU
vzD0eqVRASqAZTK2spiQsaX0pug7rWgG
fENeGhztno5lMcJDTXQfe9H6ocLZANpAF
v_p0dRbO6ggAvZN0TfyiuR4focFMAjr6VF
GVbprEQOEaS1KRs8loF3FgL0rsmP81K8
06Gv5zeSurK_sp8GEelqtm1G3J059zUdP
aJz7zJQEFPSH0TXv3lml5u9lnvnAbM0jCl
F03nG5ZjB-6WUPfS41Os5DgzTpSzUTR
-YZc1HnRUxy6l5grDPFmDiNZn0Ie9Mrlkc
PI2A8dubQZm3d8pYVpg6Y2OqzifmZzNs
QM.Zuh5Ko4bf20Bz46cNFRHJvcyPfJyglp
cvN1uM16-yOI&dib_tag=se&keywords=m4
+viti+testa+svasata&nsdOptOutParam=tru
e&qid=1734712054&s=industrial&sprefix=
m4+viti+testa+svasata%2Cindustrial%2C1
28&sr=1-6		M4x12 - Flat head
ScrewsGlobstarhttps://www.amazon.it/testa-svasata-
impronta-Phillips-acciaio/dp/B083RH
5JYL/ref=sr_1_9?__mk_it_IT=%C3%
85M%C3%85%C5%BD%C3%95%C
3%91&crid=XEVGIE9R1K2F&dib=ey
J2IjoiMSJ9.sYNfljBDPU1N2px8nueT
zi6D61Z46HD98MSuIwqXNKrfkSOM
GYgywNTC8-dsYqNXiZEMS42kGB4
QdBtz_TVFLvgLCFQVl9W5n7Bo4fQS
_VKHh9VLmA_RjdF75wPsSbRPIdSA
nidi0pehfkRoBIrEtYVn-cUq4TXH0P1D
pGLc_4J88tU02ieLVee-YarMxZLKeLYt
kK6B2CmM6d_AE0K7UJofwnOCOLp
3R72KpMTltjkJutf0WxBFsNQhYG8Wz
0q1o_3uRs_aNM0wJtGeSohS3fABN8C
z-kYxdfOikF1zi4nE1wn8YUw8lfsCxeUI
ioUp4Lz0GaXhHkkXxC5jssOsuOyW_6
CeDYrx0AOAy1URHBY.4xc_PNzUrZe
FFz0FV9hV7uu_-Es6f3jhi6koImPVcgw
&dib_tag=se&keywords=m4x70%2Bvi
ti%2Btesta%2Bsvasata&nsdOptOutPa
ram=true&qid=1734712182&s=industri
al&sprefix=m4x70%2Bviti%2Btesta%2
Bsvasata%2Cindustrial%2C134&sr=1-9&th=1		M4x70 - Flat head
ScrewsAerzetixhttps://www.amazon.it/AERZETIX-cilin
drica-impronta-intaglio-montaggio/dp/B
0BLNJ9PTX/ref=sr_1_8?__mk_it_IT=%
C3%85M%C3%85%C5%BD%C3%95
%C3%91&crid=3AESCF6VKG6B6&di
b=eyJ2IjoiMSJ9.3HlweLtFpKk8huuM4
tfcAbDSl2biH6iWGO7PXpXKxzQnVM
cKVrvCI0CkZanzuaduqvaeCs8DLxlFs
c_s4-qvSDCE8dvXDhuO4nHPwIhPAl
BnL5G6tLMycq0ZS0ZN83GAj_VOUc
GVLq4fdcfeurabYdPzrWf3PpwM6_-y
OlEOLVvlzzBKgQWO7fiMSCFuGA6
X4j8xVlNzobgrSfsnDNqZpEgH289rp
AslvAA9D2YEmvJSm0CRqWecsKPJ
iB8yNhelz01McT-qy3s6utg4_Thec18s
iTwIY2bFdQ5XYhkTzBxAF86lr6VMXZ
3Ym26ZiedQE1_7ceB7jYcyQ5mOMR
fx0d8pYVpg6Y2OqzifmZzNsQM._rEh
uS8xCOyhS7xqQQe9H3kwn-ppl22Rf
VmzFpFpyW0&dib_tag=se&keyword
s=m4x80%2Bviti%2Btesta%2Bsvasat
a&nsdOptOutParam=true&qid=17347
12214&s=industrial&sprefix=m4x80%
2Bviti%2Btesta%2Bsvasata%2Cindu
strial%2C141&sr=1-8&th=1		M4x80 - Flat head
ScrewsGlobstarhttps://www.amazon.it/testa-svasata-
impronta-Phillips-acciaio/dp/B083RJ
S9NT/ref=sr_1_30?__mk_it_IT=%C3
%85M%C3%85%C5%BD%C3%95%
C3%91&crid=15MBBIPYCF06S&dib
=eyJ2IjoiMSJ9.wHpVBrZH8GsdSIko
N0T0-2663KN2Tr-H4orSaQtmYf64T
nTN15zshBEbfwlrPNyJaD4F1cXZvO
Evle2B5qQb_Un-tR8WzVwAxQFWe
SMPaYKjM5ZgpbhFaaS6Zj0BQntEIA
hyTnvnLg4IBEE9y-4jQ66ZIkmXcWFo
zXK-eTFnle6uFI4nXhbCXTWjM5MP_
sNolDP0O4HS7Bnxk_wanuwC9zEuH
ZeVNsejl6gwYR0IlJK5iu0V9Y-bgFcr1-
iTBhrARDKG7-0Nc5188TSaN9snT7K
6yO2J3ctfC24PTGnaZAFYKMvrNqz
OAihVBKIW8xOhIx0ZRipPa9ggsMs
_pe1VgY2dCXtUrE_bo1dd4oGlUuw
.v5iiMZ2rMQlZlZs2WDyqM_mo1utvu
PD71Jyb-O7BY1Y&dib_tag=se&key
words=m5x8+viti+testa+svasata&ns
dOptOutParam=true&qid=17347123
79&s=industrial&sprefix=m5x8+viti+
testa+svasata%2Cindustrial%2C201
&sr=1-30		M5x8 - Flat head
ScrewsGlobstarhttps://www.amazon.it/testa-svasata-
impronta-Phillips-acciaio/dp/B083RH
M4BY/ref=sr_1_30?__mk_it_IT=%C3
%85M%C3%85%C5%BD%C3%95%
C3%91&crid=15MBBIPYCF06S&dib=
eyJ2IjoiMSJ9.wHpVBrZH8GsdSIkoN
0T0-2663KN2Tr-H4orSaQtmYf64Tn
TN15zshBEbfwlrPNyJaD4F1cXZvO
Evle2B5qQb_Un-tR8WzVwAxQFW
eSMPaYKjM5ZgpbhFaaS6Zj0BQnt
EIAhyTnvnLg4IBEE9y-4jQ66ZIkmX
cWFozXK-eTFnle6uFI4nXhbCXTW
jM5MP_sNolDP0O4HS7Bnxk_wan
uwC9zEuHZeVNsejl6gwYR0IlJK5iu
0V9Y-bgFcr1-iTBhrARDKG7-0Nc5
188TSaN9snT7K6yO2J3ctfC24PTG
naZAFYKMvrNqzOAihVBKIW8xOhI
x0ZRipPa9ggsMs_pe1VgY2dCXtUr
E_bo1dd4oGlUuw.v5iiMZ2rMQlZlZs
2WDyqM_mo1utvuPD71Jyb-O7BY
1Y&dib_tag=se&keywords=m5x8%
2Bviti%2Btesta%2Bsvasata&nsdOpt
OutParam=true&qid=1734712379&s
=industrial&sprefix=m5x8%2Bviti%2
Btesta%2Bsvasata%2Cindustrial%2
C201&sr=1-30&th=1		M5x10 - Flat head
ScrewsGlobstarhttps://www.amazon.it/testa-svasata-
impronta-Phillips-acciaio/dp/B083RH
9SYZ/ref=sr_1_30?__mk_it_IT=%C3
%85M%C3%85%C5%BD%C3%95%
C3%91&crid=15MBBIPYCF06S&dib
=eyJ2IjoiMSJ9.wHpVBrZH8GsdSIko
N0T0-2663KN2Tr-H4orSaQtmYf64T
nTN15zshBEbfwlrPNyJaD4F1cXZv
OEvle2B5qQb_Un-tR8WzVwAxQF
WeSMPaYKjM5ZgpbhFaaS6Zj0BQ
ntEIAhyTnvnLg4IBEE9y-4jQ66ZIkm
XcWFozXK-eTFnle6uFI4nXhbCXT
WjM5MP_sNolDP0O4HS7Bnxk_w
anuwC9zEuHZeVNsejl6gwYR0IlJK5
iu0V9Y-bgFcr1-iTBhrARDKG7-0Nc5
188TSaN9snT7K6yO2J3ctfC24PTG
naZAFYKMvrNqzOAihVBKIW8xOhIx
0ZRipPa9ggsMs_pe1VgY2dCXtUrE
_bo1dd4oGlUuw.v5iiMZ2rMQlZlZs2
WDyqM_mo1utvuPD71Jyb-O7BY1Y
&dib_tag=se&keywords=m5x8%2Bvi
ti%2Btesta%2Bsvasata&nsdOptOutP
aram=true&qid=1734712379&s=indus
trial&sprefix=m5x8%2Bviti%2Btesta%
2Bsvasata%2Cindustrial%2C201&sr=
1-30&th=1		M6x20 - Flat head
SD cardSanDiskhttps://www.amazon.it/SanDisk-Ultra-
Scheda-memoria-classe/dp/B08GYG6
T12/ref=sr_1_8?crid=3ETH6HJL8VKD
A&dib=eyJ2IjoiMSJ9.5omqicizrizfHo8r
pPQ6sWJ_C0sfwUqPQv6VrziwK-SCX
4-aSgFGois1pfvTwB0SuDxdNjByo2zU
fBonWwUKAf3clazXEIGNl0GhojQmvj
Bt_6RS31QcAM5Y50BpAdndMXIHy9
iGTswgYEwnbt9uj56G_soC6OdPiTvr
W3M8jwtTxQDUl8UcHZeRMPq0T8sd
v4hskHGmipFItAzE1lyDiy0otrqUhz-ph
OggXBAM6gMyGf0qBtU1AFwfj81Shl
HS2qojCiRq_JLKyBkc_Y4P_V1YtsGc
MxnoG6YzYowwEYbX7uTFWJB-Wvc
B_vYw-D5s0B-HDSYRSl-C60yhtqhrUw
grg9w6PN5U5s01vos4bauIgIN6lOez1
tz2OHwL5YL3QCHKxNnJV0IM42cH0
du8fIEIbUwYC5jRLqowx1UyaQKQ4W
WaGozdZ9l5qUoeQ7KU.iOPo2BYLK
STjqjB8BevaVgPa5QD7BuP9CieN6u
4_Qeo&dib_tag=se&keywords=sd%
2Bcard%2Bsandisk&nsdOptOutParam
=true&qid=1734704871&sprefix=SD%
2Bcard%2Bsan%2Caps%2C141&sr=8-8&th=1		SanDisk 32 GB Extreme PRO SD card, UHS-I
Self-aligning Flange bearingBrateuanoiiKFL088 mm inner diameter
Slicing softwareUltimaker Curahttps://ultimaker.com/it/software/
ultimaker-cura/?gad_source=1&
gclid=CjwKCAiAyJS7BhBiEiwAy
S9uNUjnbkNIYNUco6YaZYHcFT
2tqBac23MU4Qx3FaMXjl_UifJ_
k8as3BoCeVkQAvD_BwE		Ultimaker Cura 4.9.0
SpringsYIXISIhttps://www.amazon.it/sspa/click?ie
=UTF8&spc=MTo4NDAzNDQ1MzM
0OTUyODQzOjE3MzQ3MDQ4MD
c6c3BfYXRmOjMwMDM5MDQzMz
kzODAzMjo6MDo6&url=%2FYIXIS
I-Assortimento-Estensione-Compre
ssione-Dispositivi%2Fdp%2FB0DD
T3KTBF%2Fref%3Dsr_1_1_sspa%
3F__mk_it_IT%3D%25C3%2585M
%25C3%2585%25C5%25BD%25
C3%2595%25C3%2591%26crid%
3D1NT7CA1YJGWK4%26dib%3D
eyJ2IjoiMSJ9.IhFmevq0fXTGGwE
cGvfUxxLOudQqQcPjSGXlWklXO
HY2Ahr2XxWSg6MaNBgZ71N1pQ
h9Wkp5c2ary4Hm2xVi4dTH8Ny4
4-OhlcrTn0G8emvX66Zu7U0LGm
xGYKyKCGxzlhwYC4EnlmTSH_O
JnA5oG-BQzuxFBdCrce0txErCEm
S40xONaaYp2R5yMiPDw2PqsPP
jLmoSCYs4XctxEvAwn_5rp71t0U
QsZNku6-jDiiuh5klpC9DsnjipLFP
unRCbITWnw6wslENLdJtPDOGF
8vbO5WFf7PnUy4BZ7tUPt7I.B6j
eUKwx1_t0sPaxmSHYVbRd1MG
w-kosEYkHuKlPtpQ%26dib_tag%
3Dse%26keywords%3Dmolle%2B
YIXISI%26nsdOptOutParam%3Dtr
ue%26qid%3D1734704807%26spr
efix%3Dmolle%2Byixisi%252Caps
%252C137%26sr%3D8-1-spons%2
6sp_csd%3Dd2lkZ2V0TmFtZT1zcF
9hdGY%26psc%3D1
Stepper motorTwotrees 123417HS4401Stepper motor bipolar NEMA 17 42 Ncm 1.5 A
Stepper motorSTEPPERONLINEUL1007AWG6Stepper motor bipolar NEMA 17 42 Ncm 1.5 A
Stepper motor fixing bracketYOTINOhttps://www.amazon.it/gp/product/
B07D7XH97T/ref=ppx_yo_dt_b_
search_asin_title?ie=UTF8&psc=1
Threaded nut M2RS-components281-984
Threaded nut M3RS-components281-984
Threaded nut M4RS-components281-984
Threaded nut M5RS-components281-984
Threaded nut M6RS-components281-984
TRAMP - C1 cell lineATCCCRL-2730Epithelial cell line isolated from the prostate of an adult male transgenic mouse with adenocarcinoma
Trypan blueGibco Thermofisher15250061
TrypsinGibco Thermofisher15400054Sterile

References

  1. Patel, T. et al. Multicellular tumor spheroids: A convenient in vitro model for translational cancer research. Life Sci. 358, 123184 (2024).
  2. Tanaka, M. et al. Emerging translational research in neurological and psychiatric diseases: From in vitro to in vivo models. Int J Mol Sci. 24 (21), 15739 (2023).
  3. De Maria Marchiano, R. et al. Translational research in the era of precision medicine: Where we are and where we Will Go. J Pers Med. 11 (3), 216 (2021).
  4. Knowlton, S. et al. Bioprinting for cancer research. Trends Biotechnol. 33 (9), 504-513 (2015).

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