Protocols
Stemgent® protocols

Thawing StemRNA hiPSCs

Thawing, Passaging, Culturing, and Cryopreservation of Human MSCs

Detection of AP using the Alkaline Phosphatase Staining Kit II

RNA Transfection using the Stemfect RNA Transfection Kit

hPSC Culture using NutriStem hPSC XF Culture Medium

Cryopreservation of human ES cells using CryoStem Freezing Medium

Using the hES Cell Cloning and Recovery Supplement to Enhance Thawing and Recovery, Single Cell Cloning, and Passaging of hES Cells

Immunofluorescence Staining of Cells for Flow Cytometry

General Protocol for Immunocytochemistry

General Protocol for ICC Staining of Live Cells

Stemgent® StemRNA™ 3rd Gen Reprogramming Kit for Reprogramming Adult and Neonatal Human Fibroblasts

Stemgent StemRNA 3rd Gen Reprogramming Kit for Reprogramming Endothelial Progenitor Cells (EPCs) (From Blood)

Stemgent StemRNA 3rd Gen Reprogramming Kit for Reprogramming Urine-Derived Progenitor Cells (UPCs)

iMatrix-511 Cell Culture Substrate: Coating Culture Vessels

General Protocol for Western Blot Analysis
Alvetex® protocols
Alvetex® Scaffold
1. Basics

Alvetex Scaffold: quick start

Media volumes guide for different feeding options in Alvetex Scaffold in well insert formats

Alvetex Scaffold 96 well plate: quick start

Alvetex Scaffold 96 well plate: instructions for use and example applications

Alvetex Scaffold 384 well plate: quick start

Alvetex Scaffold 384 well plate: instructions for use and example application
3. Cell-Specific

Guidance notes and protocols for the culture of primary rat hepatocytes in Alvetex Scaffold in well insert and well plate formats

Protocol for the culture of primary rat hepatocytes in Alvetex Scaffold 96 well plate format

Culture of the canine kidney epithelial MDCK cell line on Alvetex Scaffold in well insert and well plate formats

Example protocol for the culture of the MCF-10A cell line on Alvetex Scaffold (12 well insert format and 96-well plate format)

Example protocol for the culture of the CaCo2 cell line on Alvetex Scaffold in well insert and well plate formats with or without a collagen I gel layer

Example protocol for the co-culture of the Caco-2 cell line with the CCD-18co cell line on Alvetex Scaffold in well insert formats

Example protocol for the culture of the breast carcinoma MDA-MB-231 cell line on Alvetex Scaffold in well insert and well plate formats

Example protocol for the culture of the human colon fibroblastic CCD-18co cell line on Alvetex Scaffold in well insert and well plate formats

Example protocol for the culture of the lung carcinoma A549 cell line on Alvetex Scaffold in well insert and well plate formats

Example protocol for the culture of the breast invasive ductal carcinoma BT-474 cell line on Alvetex Scaffold

Example protocol for the culture of the PC-3 cell line on Alvetex Scaffold in well plate and well insert formats

Example protocol for the culture of the LN-229 cell line on Alvetex Scaffold in 96 well plate format

Example protocol for the culture of the 3T3 cell line on Alvetex Scaffold in well plate and well insert formats

Example protocol for the culture of the TERA2.cl.SP12 cell line on Alvetex Scaffold

Example protocol for the culture of the HepG2 cell line on Alvetex Scaffold in well plate and well insert formats

Example protocols for the culture of the CHO-K1 cell line on Alvetex Scaffold in well plate and well insert formats

Example preparation of HaCaT epidermal equivalent using Alvetex Scaffold

Bioengineering of human full thickness skin for in vitro applications, screening and product development

Example protocol for the culture of the SW480 cell line on Alvetex Scaffold in well insert and well plate formats

Example protocol for the culture of the SW620 cell line on Alvetex Scaffold in well insert and well plate formats

Example protocols for the co-culture of either the SW620 or the SW480 cell line with the 3T3 cell line on Alvetex Scaffold in well insert formats

Example protocol for the culture of the MG63 cell line on Alvetex Scaffold in well insert formats

Example protocol for the culture of primary rat MSCs on Alvetex Scaffold in well insert formats

Example protocols for the culture of the MCF-7 cell line on Alvetex Scaffold in well plate and well insert formats
4. Transfection

3D Transfection System (Mirus)
5. Cell Visualization

Simple visualisation of cells on Alvetex Scaffold using light microscopy (Neutral Red-Methyl Blue dye)

Live cell imaging of 3D cultures grown on Alvetex Scaffold using confocal microscopy

Wheat germ agglutinin staining of 3D cultures grown on Alvetex Scaffold and Alvetex Strata
6. Metabolic Assays

alamarBlue® cell viability assay of upcyte® hepatocyte cultures grown on Alvetex Scaffold in 3D

Determination of cell number and growth characteristics within Alvetex Scaffold using the PicoGreen® assay

XTT cell viability assay for Alvetex Scaffold 96 well plate format

MTS cell viability assay of upcyte® hepatocyte cultures grown on Alvetex Scaffold in 3D

MTT cell viability assay of cultures grown on Alvetex Scaffold in 3D
7. Genomics and Proteomics

Total protein extraction from cells cultured in Alvetex Scaffold in 3D

Extraction of RNA from cells cultured in Alvetex Scaffold in 3D

Protein extraction from cells cultured in Alvetex Scaffold 96 well plate format

Protein extraction from cells cultured in Alvetex Scaffold 96 well plate format
8. Histology

Histology (1): Choosing the Right Fixative to Preserve 3D Cell Cultures

Histology (2): Processing Alvetex Scaffold 3D Cultures for PFA Fixation and Paraffin Wax Embedding for Immunofluorescence

Histology (3): Processing Alvetex Scaffold 3D Cultures for Bouin’s Fixation and Paraffin Wax Embedding for Haematoxylin and Eosin Staining

Histology (4): Processing Alvetex Scaffold 3D Cultures for Resin Embedding and Toluidine Blue Staining for Light Microscopy

Histology (5): Processing Alvetex Scaffold 3D Cultures for Scanning Electron Microscopy (SEM)
11. 3D Perfused Cell Culture

Example Protocol using Alvetex™ Scaffold 6 Well Inserts for 3D Perfused Cell Culture (Alvetex Perfusion Plate)
REPROCELL® protocols

Cell Cultivation Protocol for ReproHepato™ (Cat.# RCDH001N)

Calcium imaging of ReproNeuro™ (Cat.# RCESDN001, RCESDN002, RCDN002N, RCDN003P)

Calcium imaging of ReproNeuro™ cells (Cat.# RCDN002N, RCDN003P, RCDN001N)

Protocol of ReproNeuro™ (Cat.# RCDN002N, RCDN003P)

Protocol of ReproNeuro™ (Cat.# RCDN002N, RCDN003P, RCDN001N)

ReproCardio 2™ (Cat.# RCDC001N)

Cryopreservation with ReproCryo DMSO Free™ (Cat.# RCHEFM002)

Feeder Cells (MEF) (Cat.# RCHEFC003)

Feeder Cells (SL10) (Cat.# RCHEFC001)

Freezing Medium for human ES/iPS Cells (Cat.# RCHEFM001)

Primate ES Cell Medium (Cat.# RCHEMD001)

ReproFF2™ (Cat.# RCHEMD006)

Dissociation Solution for human ES/iPS Cells (Cat.# RCHETP002)

ReproMed™ iPSC Medium (Cat.# RCRM101)

Primary Hepatocyte culture with ReproHP Medium™ (Cat. No. RCRM201 (and RCRM201-S))
Protocols from other brands
ABLE® Biott®

Operating instructions for ABLE Biott Bioreactor System