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CAS Trypsin: 9002-07-7 / EDTA: 60-00-4 Cell Dissociation Reagent Cell Dissociation Reagent

Trypsin-EDTA Solution

IUPAC: N/A (formulated enzyme-chelator mixture)
CAS NUMBER
Trypsin: 9002-07-7 / EDTA: 60-00-4
MOL. FORMULA
N/A (formulated mixture)
MOL. WEIGHT
N/A
Scientific Overview

Trypsin cleaves cell-surface and extracellular-matrix adhesion proteins that anchor adherent cells to the culture substrate, while EDTA chelates divalent cations (Ca2+/Mg2+) required for cadherin-mediated cell-cell and cell-substrate adhesion, synergistically enhancing and accelerating cell detachment compared to trypsin alone. Because residual trypsin activity can damage cell-surface receptors if exposure is prolonged, the reaction is typically neutralized promptly with serum-containing medium or a trypsin inhibitor after cells detach.

Trypsin-EDTA Solution is a ready-to-use cell dissociation reagent combining the serine protease trypsin with the calcium/magnesium-chelating agent EDTA, used routinely to detach adherent mammalian cells from culture vessel surfaces for passaging, harvesting, or downstream analysis.

Full Technical Data
Chemical Name Trypsin-EDTA Solution (0.25% Trypsin / 0.02-0.53 mM EDTA)
CAS Number Trypsin: 9002-07-7 / EDTA: 60-00-4
IUPAC Name N/A (formulated enzyme-chelator mixture)
Molecular Formula N/A (formulated mixture)
Molecular Weight N/A g/mol
Reaction Scheme
Porcine Pancreas (Trypsinogen) + EDTA (Disodium Salt)
NA
NA
Trypsin-EDTA Solution
NA
Commercial Applications

Adherent Cell Passaging

Standard reagent for routine subculturing/passaging of adherent mammalian cell lines in research and bioprocessing.

Cell Harvesting for Downstream Analysis

Used to detach cells intact for flow cytometry, cell counting, and other single-cell analytical workflows.

Primary Cell Isolation

Assists in dissociating tissue explants and primary cell cultures into single-cell suspensions.

Cell & Gene Therapy Manufacturing

Used in scale-up cell expansion and harvest steps within GMP cell-therapy manufacturing workflows.

Market Therapeutic Focus
CELL DISSOCIATION REAGENT PHARMACEUTICAL MANUFACTURING CELL & GENE THERAPY / BIOPROCESSING ACADEMIC & INDUSTRIAL RESEARCH DIAGNOSTICS & IMMUNOASSAYS CRO/CDMO
Scientific Literature

Optimization of Trypsinization Protocols for Mammalian Cell Culture

Authors: Freshney, R.I.

Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications, Wiley

Effects of Trypsin/EDTA Treatment on the Expression of Cell Surface Markers

Authors: Grinnell, F.

Exp. Cell Res.

Cell Detachment Methods for Adherent Cell Culture: A Comparative Review

Authors: Nagata, S. et al.

J. Biotechnol.

Peer Queries & FAQs
Why combine EDTA with trypsin instead of using trypsin alone?
EDTA chelates the calcium and magnesium ions that stabilize cell-cell and cell-matrix adhesion proteins, weakening those bonds and allowing trypsin to detach cells faster and more gently.
How is the trypsinization reaction stopped?
It is typically neutralized by adding serum-containing complete growth medium (serum proteins competitively inhibit trypsin) or a defined trypsin inhibitor for serum-free workflows.
Can prolonged trypsin exposure damage cells?
Yes, excessive exposure can cleave cell-surface receptors and reduce viability, so cells should be monitored and the reaction neutralized as soon as detachment is observed.
What is the recommended storage condition?
Store frozen at -10 to -30°C for long-term storage, or at 2-8°C for short-term use; avoid repeated freeze-thaw cycles.

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