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Cellworks

Welcome to Cellworks - A division of Caltag Medsystems

Cellworks, a division of Caltag Medsystems, specialises in the manufacture and development of Angiogenesis focused cells, kits and reagents for in vitro co-culture angiogenesis assays. Angiogenesis assays can be measured dynamically or by end point staining.Launched in November 2019, Cellworks now provide the AngioCo Angiogenesis Co-culture Assay 96wp Kit (GFP-HUVEC/HDF), a validated, in vitro, human primary cell co -culture model of angiogenesis that allows the dynamic measurement of all phases of the angiogenic process in a 96 well plate format.

Introduction to Angiogenesis

Angiogenesis is the development of new capillaries from existing vasculature and plays important roles in embryogenesis, the female reproductive cycles, bone formation and wound healing. However, excessive angiogenesis can cause tumour development, diabetic retinopathy and psoriasis.

AngioCo Angiogenesis Co-culture Assay Kit (GFP-HUVEC/HDF)

AngioCo

Product Codes: ZHA-4100-24 and ZHA-4100-96

The AngioCo Angiogenesis Co-culture Assay Kit (GFP-HUVEC/HDF) is a validated, in vitro, human primary cell co -culture model of angiogenesis that allows the dynamic measurement of all phases of the angiogenic process.

Tubules formed contain a lumen and resemble the morphology and range of capillary lengths found in microvascular beds in vivo.

Formerly known as GFP V2a Kit - Vasculogenesis to Angiogenesis, these are new versions of the kit with the following improved features:

  • Now available in two formats: 24 or 96 well plate. The new 96 well plate allows for a greater number of tests to be ran in unison
  • Single vial of co-culture cells containing HUVECs and Fibroblasts
  • Assay length has been reduced from 14 days to 11 days, allowing for quicker results.
  • Now able to seed and grow cells in the same medium
  • The tubule formation has been optimised for the control wells
  • There is a greater difference between Suramin and VEGF controls, allowing for easier visualistaion
  • Now shipped in less packaging

The AngioCo Angiogenesis Co-culture Assay Kit (GFP-HUVEC/HDF) enables the dynamic measurement of vascular tube formation.

  • It is a kinetic, quantitative, in both 24 well and 96 well format, co culture angiogenesis model,utilising lentivirally infected human umbilical vein endothelial cells (HUVEC) expressing GFP co-cultured with normal human dermal fibroblasts (HDF).
  • This co-culture model recapitulates all phases of the in vivo process from vasculogenesis to angiogenesis (including: cell activation, cell proliferation, migration, elongation and anastomosis)
  • Test compounds, media or tissue explants can be added at any point in the pathway - from the onset of vasculogenesis through to advanced angiogenesis - giving total assay flexibility
  • The AngioCo Angiogenesis Co-culture Assay Kit (GFP-HUVEC/HDF) includes: +/- controls, media, supplements and a 24 or 96 well plate
  • AngioCo Co-Culture Cells are provided frozen allowing long term storage and user determined assay initiation
  • Tubule growth is quantifiable using AngioSys 2.0 Software
Tubule Formation in response to VEGF

Tubule Formation in response to VEGF

Untreated Tubules

Untreated Tubules

Tubule Formation in response to Suramin

Tubule Formation in response to Suramin

AngioCo Angiogenesis Co-culture Assay Kit (GFP-HUVEC/HDF)

Product Codes: ZHA-4100-24 and ZHA-4100-96

Kit Contents:

  • Ampoule of cryopreserved AngioCoCo-Culture Cells - (GFP-HUVEC/HDF)
  • 24 or 96 well tissue culture plate
  • 125ml Growth Medium
  • Angiogenesis Control compounds (VEGF and Suramin)
  • Growth Supplement
  • Antibiotic/Antimyotic

For storage conditions and additional equipment required please read the relevant protocol:

Molecules shown to be active in the Cellworks AngioCo Angiogenesis Co-Culture Assays Inhibitors

Inhibitors

VEGF neutralising antibody, VEGF antisense oligonucleotide, Collagen IV neutralising antibody, PECAM-1 neutralising antibody, VEGF-R antagonists, Thrombospondin derivatives, TNF alpha, TGF b1, aVb3 integrin antagonists, Suramin, Curcumin, Fumagillin, TNP-740 (fumagillin analogue), Colchicine, Cytochalasin B, Erucic acid, Ginsengosides, RGD peptide, Endostatin, Thymine, Thalidomide and analogues, PAI-1 inhibitors MMP inhibitors, MGBG (methylglyoxalbisguanylhydrozone), some conditioned media.

Enhancers

VEGF, PIDGF, BFGF, 2-deoxyribose-1-P, Most tumour cell conditioned media, Wound fluid, Ascitic fluid from leiomyosarcoma patient, Ascorbic acid (biphasic response).

Note: that this represents only the molecules whose identity we know. We have tested many proprietary compounds from various laboratories that were active in both senses but which were supplied as coded samples whose structure is not known to us.

Quantification of In Vitro Angiogenesis through Image Analysis

Cellworks offer two possibilities for analysis of in vitro angiogenesis assays based on the co-culture of endothelial cells with fibroblasts. Both provide accurate quantification of the stimulatory and inhibitory effects of different agents and output the lengths of the tubule complexes as well as the numbers of junctions in each.

V2a Kit - Vasculogenesis to Angiogenesis

Some publications make reference to the "GFP V2a Kit - Vasculogenesis to Angiogenesis". This has been superseded by the AngioCo Angiogenesis Co-culture Assay due to customer feedback.

AngioCo Angiogenesis Co-culture Assay 24wp Kit (HUVEC/HDF)

AngioCo

'An in vitro human primary cell co-culture model of angiogenesis that includes all the phases of the angiogenic process'

Tubules formed contain a lumen and resemble the morphology and range of capillary lengths found in microvascular beds in vivo.

Formerly known as V2a Kit - Vasculogenesis to Angiogenesis, this is a new version of the kit with the following improved features:

  • Assay length has been reduced from 14 days to 11 days, allowing for quicker results
  • Single vial of co-culture cells containing HUVECs and Fibroblasts
  • Now able to seed and grow cells in the same medium
  • The tubule formation has been optimised for the control wells
  • There is a greater difference between Suramin and VEGF controls, allowing for easier visualistaion
  • Now shipped in less packaging

Benefits

  • Analyse the growth pathway from vasculogenesis to angiogenesis (including: cell activation, cell proliferation, migration, elongation and anastomosis)
  • Add test compounds, media or tissue explants at any point in the pathway - from the onset of vasculogenesis continuing through to advanced angiogenesis
  • Intervene in the cellular growth process at any stage
  • Kit includes: +/- controls, media, supplements, staining components, plate
  • Frozen cells: perform assay at your convenience
  • Large batches of cells in stock
  • Tubule growth is quantifiable using AngioSys 2.0 Software

Tubule Formation in response to VEGF
Tubule Formation in response to VEGF

Untreated Tubules
Untreated Tubules

Tubule Formation in response to Suramin
Tubule Formation in response to Suramin

AngioCo Angiogenesis Co-culture Assay 24wp Kit (HUVEC/HDF)
   Product Code: ZHA-4000-24

Kit Contents:

  • Ampoule of cryopreserved AngioCoHUVEC/HDF Cells
  • 24 well tissue culture plate
  • Angiogenesis Growth Medium
  • Growth Supplement
  • Angiogenesis Control compounds (VEGF and Suramin)
  • Mouse anti-human CD31 Primary Antibody
  • Goat anti-mouse IgG Alkaline Phosphatase Secondary Antibody
  • BCIP/NBT insoluble substrate (for permanent record of tubule staining)

For storage conditions and additional equipment required please read the protocol:

Molecules shown to be active in the Cellworks AngioCo Angiogenesis Co-Culture Assay:

Inhibitors

VEGF neutralising antibody, VEGF antisense oligonucleotide, Collagen IV neutralising antibody, PECAM-1 neutralising antibody, VEGF-R antagonists, Thrombospondin derivatives, TNF alpha, TGF b1, aVb3 integrin antagonists, Suramin, Curcumin, Fumagillin, TNP-740 (fumagillin analogue), Colchicine, Cytochalasin B, Erucic acid, Ginsengosides, RGD peptide, Endostatin, Thymine, Thalidomide and analogues, PAI-1 inhibitors MMP inhibitors, MGBG (methylglyoxalbis(guanylhydrozone), some conditioned media.

Enhancers

VEGF, PIDGF, BFGF, 2-deoxyribose-1-P, Most tumour cell conditioned media, Wound fluid, Ascitic fluid from leiomyosarcoma patient, Ascorbic acid (biphasic response).

Note: that this represents only the molecules whose identity we know. We have tested many proprietary compounds from various laboratories that were active in both senses but which were supplied as coded samples whose structure is not known to us.

Quantification of In Vitro Angiogenesis through Image Analysis

Cellworks offer two possibilities for analysis of in vitro angiogenesis assays based on the co-culture of endothelial cells with fibroblasts. Both provide accurate quantification of the stimulatory and inhibitory effects of different agents and output the lengths of the tubule complexes as well as the numbers of junctions in each.

V2a Kit - Vasculogenesis to Angiogenesis

Some publications make reference to the "AngioKit" and "V2a Kit - Vasculogenesis to Angiogenesis". These have both been superseded by the AngioCo Angiogenesis Co-culture Assay due to customer feedback.

Build a Bespoke Angiogenesis Assay

'Cellworks provide angiogenesis validated cells and media for maximum assay flexibility. Both GFP human umbilical vein endothelial cells (HUVEC) and normal HUVEC are available, allowing dynamic measurements of angiogenesis to be taken or end point staining.'

Bespoke assays allow the user to have complete control over the number and cell types present in the angiogenesis assay and the opportunity to manipulate cells prior to culturing if necessary.

Benefits

  • Purchasing components separately allows total flexibility in terms of cell storage, assay set up and test agent introduction
  • Control over the number and types of cells that can be used in the assay
  • Cells can be manipulated prior to culturing, lending the assay to more detailed studies
  • Cells have proven angiogenesis potential (tubule formation demonstrated using the Cellworks -AngioCo Angiogenesis Co-Culture assay as part of the QC process for all batches of cells)
  • Tubule growth can be analysed using the Cellworks AngioSys software

Angiogenesis Visualisation of tubule formation through GFP transfection of HUVEC cells

Angiogenesis Fibroblasts

Angiogenesis Tubules stained with anti CD31 (PECAM-1)

GFP Human Umbilical Vein Endothelial Cells (HUVEC) (angiogenesis co-culture validated)
   Product Code: ZHC-2402
  • Produce tubules in conjunction with Cellworks Human Dermal Fibroblasts (HDF) - Angiogenesis Co-Culture Validated (ZHC-5102), when cultured with Cellworks Angiogenesis Growth Medium (ZHA-1960 and ZHA-1970).
  • Validated for use in the manufacture of Cellworks AngioCo Angiogenesis Co-Culture Assay Kit (GFP-HUVEC/HDF) (Product Codes: ZHA-4100-24 and ZHA-4100-96).
  • Minimum of 500,000 cryopreserved cells.
  • Guaranteed for 10 population doublings.
Human Umbilical Vein Endothelial Cells (HUVEC) (angiogenesis co-culture validated)
   Product Code: ZHC-2102
  • Produce tubules in conjunction with Cellworks Human Dermal Fibroblasts (HDF) - Angiogenesis Co-Culture Validated (ZHC-5102), when cultured with Cellworks Angiogenesis Growth Medium (ZHA-1970).
  • Validated for use in the manufacture of Cellworks AngioCo Angiogenesis Co-Culture Assay Kit (GFP-HUVEC/HDF) (Product Codes: ZHA-4100-24 and ZHA-4100-96).
  • Minimum of 500,000 cryopreserved cells.
  • Guaranteed for 10 population doublings.
Human Dermal Fibroblasts (HDF) - Angiogenesis Co-Culture Validated
   Product Code: ZHC-5102
  • Produce tubules in conjunction with Cellworks Human Umbilical Vein Endothelial Cells (HUVEC) (angiogenesis co-culture validated (ZHC-2102 and ZHC-2402), with Cellworks Angiogenesis Growth Medium (ZHA-1970).
  • Validated for use in the manufacture of Cellworks AngioCo Angiogenesis Co-Culture Assay Kit (GFP-HUVEC/HDF) (Product Codes: ZHA-4100-24 and ZHA-4100-96).
  • Minimum of 1,000,000 cryopreserved cells.
  • Guaranteed for 10 population doublings.
Early Passage Human Umbilical Vein Endothelial Cells (HUVEC)
   Product Code: ZHC-2301
  • Early Passage HUVECs are cryopreserved at the end of the secondary culture.
  • A cost effective alternative to buying primary cells.
  • Minimum of 500,000 cells.
  • Guaranteed for 10 population doublings.

Supporting Reagents

CD31 Tubule Staining Kit
Product Code: ZHA-1225
Angiogenesis Assay Control Kit
Product Code: ZHA-1300

Media and Supplements

Angiogenesis Growth Medium Package
Product Code: ZHA-1970
Human Large Vessel Endothelial Cell Growth Medium Package
Product Code: ZHM-2953
Human Fibroblast Growth Medium Package
Product Code: ZHM-5923

Quantification of In Vitro Angiogenesis through Image Analysis

Cellworks offer two possibilities for analysis of in vitro angiogenesis assays based on the co-culture of endothelial cells with fibroblasts. Both provide accurate quantification of the stimulatory and inhibitory effects of different agents and output the lengths of the tubule complexes as well as the numbers of junctions in each.

If you require a pre formed angiogenesis assay kit, rather than bespoke options, please visit the AngioCo Angiogenesis Co-Culture Assay page.

Angiogenesis Services

Compounds, media and tissue explants can be tested at any point in the angiogenic pathway using the Cellworks Angiogenesis Service. The service includes the following stages and can be tailored to your needs:

Angiogenesis Complete Assay Service

  • Performing Angiogenesis Assays
  • Capture and Analysis of Images
  • Provision of Raw Data
  • Bespoke Options

Cellworks offer two possibilities for the quantification of In Vitro Angiogenesis through Image Analysis. Both provide accurate quantification of the stimulatory and inhibitory effects of different agents and output the lengths of the tubule complexes as well as the numbers of junctions in each.

Angiosys 2.0 Image Analysis Software

  • AngioSys 2.0 Image Analysis Software (Product Code: ZHA-5000)
  • AngioSys 2.0 Image Analysis Software - Demonstration Version (Product Code: ZHA-5000D)

Angiogenesis Image Analysis Service

  • Angiogenesis Image Analysis Service - first 24 images (Product Code: ZHA-6000)
  • Angiogenesis Image Analysis Service - additional images (Product Code: ZHA-6001)
  • Angiogenesis Image Analysis Service Set Up (Product Code: ZHA-6002)

Angiogenesis Complete Assay Service

Analyse the Angiogenic Pathway: Cellworks Angiogenesis Service.

Compounds, media and tissue explants can be tested at any point in the angiogenic pathway using the Cellworks Angiogenesis Service. The service includes the following stages and can be tailored to your needs:

  1. Performing Angiogenesis Assays
  2. Capture and Analysis of Images
  3. Provision of Raw Data
  4. Bespoke Options

1) Performing Angiogenesis Assays

Using the Cellworks AngioCo Angiogenesis Co-Culture Assay Kit, Cellworks specialists offer customers the ability to outsource angiogenesis experiments.

The AngioCo Angiogenesis Co-Culture Assay Kit is in vitro human primary cell co-culture model of angiogenesis that includes all of the phases of the angiogenic process. Tubules formed contain a lumen and resemble the morphology and range of capillary lengths found in microvascular beds in vivo.

Test compounds, media or tissue explants can be added at any point in the pathway - from the onset of vasculogenesis continuing through to advanced angiogenesis (including: cell activation, cell proliferation, migration, elongation and anastomosis), allowing intervention in the cellular growth process at any stage. The Cellworks Angiogenesis Service can be carried out on any number of wells required by the customer, from a single well to multiple assays over several plates.

2) Capture and Analysis of Images

Tubule Formation in response to VEGF
Tubule Formation in response to VEGF

Untreated Tubules
Untreated Tubules

Tubule Formation in response to Suramin
Tubule Formation in response to Suramin

Angiogenesis experiments fixed and stained with anti-CD31 antibody are photographed and analysed using the Angiosys 2.0 software; a custom built software for measurement of tubule growth. Take advantage of the expertise of the in house specialists, with years of experience performing and analysing these assays at Cellworks.

3) Provision of Raw Data

Reports are presented in an excel format and contain raw data including details of the number of tubules, the number of junctions, the total tubule length, and the mean tubule length for each assay.

Report Example:

angiosys screenshot

4) Bespoke Options

The service is tailored to each project and all parts can be adjusted to your individual requirements. For further information about the service please contact cellworks@caltagmedsystems.co.uk  or call +44 (0)1280 827460.

Angiogenesis Image Analysis Software

'The AngioSys 2.0 image analysis software provides a semi-automated analysis of angiogenesis by measuring the number of tubules, the number of junctions, the total tubule length, and the mean tubule length for each image'

Angiosys 2.0 is custom built software developed to quantitate tubules formed during in vitro angiogenesis assays based on the co-culture of endothelial cells with fibroblasts.

AngioSys 2.0 is regularly used to quantitate images of angiogenesis tubules obtained from conducting the AngioCo Angiogenesis Co-Culture Assay Kit (Product Code ZHA-4000-24), which uses the anti-CD31 based staining kit (ZHA-1225).

AngioSys software can also be used in conjunction with the AngioCo Angiogenesis Co-Culture (GFP-HUVEC/HDF) Assay (Product Code: ZHA-4100-24 and ZHA-4100-96)

Image files can be conveniently grouped and processed to provide quantitative & repeatable measurements. Repetitive image processing sequences can be specified, saved, and applied to groups of images to enable rapid analysis. The resulting data is saved in a text format that can easily be read by third party packages such as Excel.

Benefits

  • Measure tubule length, number of junctions, frequency and mean number of branches
  • Operates on Windows XP, Vista, Windows 7, Windows 8, 32 bit, 64 bit
  • Able to analyse TIFF, BMP, GIF, JPEG, PNG images
  • 21 day trial available
  • Available as an internet download - quick and easy
benefits key
angiogenesis image analysis software benefits

Screenshot:

angiosys screenshot

AngioSys 2.0 Image Analysis software is available in two formats:

Angiogenesis Image Analysis Service

'Ideal for customers preferring the in-house specialists at Cellworks to analyse their images. Customer reports detail the number of tubules, the number of junctions, the total tubule length, and the mean tubule length for each image'

  • Ideal for customers not wishing to purchase the AngioSys 2.0 software - when analysis is required on only a small number of images
  • Suitable for analysis of large numbers of angiogenesis assay images - when manpower in the lab is short
  • Perfect for customers new to angiogenesis image analysis

Pricing is charged per image analysed. Please note that there is a one off set up charge payment required; product code: ZHA-6002.

Angiogenesis Image Analysis Service - first 24 images £20 per image.
   Product Code ZHA-6000
Angiogenesis Image Analysis Service - additional images £10 per image.
   Product Code ZHA-6001
Image Analysis Service Set Up £50 single payment per order.
   Product Code ZHA-6002

Report Example:

angiosys screenshot

If you are having difficulty downloading product data or require further information, please contact us.