Laminstem-521

Description

Laminstem-521 Overview

LaminStem-521 is a chemically defined, animal component-free matrix for feeder-free culture of human embryonic stem (hES) cells and induced pluripotent stem (hiPS) cells. LaminStem-521 facilitates self-renewal of both hES cells and iPS cells in a chemically defined, feeder-free and xeno-free cell culture system.

When used with NutriStem® hPSC XF Medium, LaminStem™ 521 has been proven to promote cellular survival and expansion of hES and hiPS cells after plating from single cell suspension.  When cultured with LaminStem-521, the hES and hiPS cells grow as monolayers on top of the laminin substrate and remain pluripotent without spontaneous differentiation.

Features

• Chemically defined, animal component-free matrix
• Superior to Matrigel for feeder-free culture of hES and hiPS cells using NutriStem® hPSC XF Medium
• No treatment with apoptosis inhibitors needed
• Support long-term self-renewal of human pluripotent stem cells (hPSC) and neural stem cells (NSCs)

Sample Data

Recent studies have shown that superior clonal derivation of hES cell lines is possible with the combined use of NutriStem® hPSC XF Medium and LaminStem™ 521.

Figure 1. H1 hESC were cultured with LaminStem™ 521 using NutriStem® hPSC XF Medium (day 3).

Instructions for Use

General Coating Procedure

Slowly thaw LaminStem™ 521 stock solution at 2 to 8°C.

Dilute the thawed LaminSem™ 521 with 1X DPBS containing Ca²⁺ and Mg²⁺.

Add the diluted LaminStem™521 solution to the tissue culture-treated culture dish or flask for a final coating concentration of 0.5 to 2 µg/cm².  Note that the optimal coating concentration is cell-dependent

Ensure the LaminStem™ 521 solution is coated evenly across the surface of the culture dish.

Seal the culture dish with Parafilm to prevent evaporation and allow it to sit undisturbed overnight at 2 to 8°C.  If a more rapid coating procedure is required, the coated culture dish can be incubated instead for 2 hours at 37°C.  Do not allow LaminStem™ 521 matrix to dry prior to use.

Important Notes

• If desired, the LaminStem™ 521 stock solution can be dispensed into working aliquots and stored at -20°C.
• Avoid repeated freeze/thaw cycles.
• Do not allow LaminStem™ 521 matrix to dry, as this will inactivate the solution.
• Thawed LaminStem™ 521 stock solution is stable for 3 months when stored under aseptic conditions at 2°C to 8°C.
• For convenience, sealed coated plates and diluted coating solution can be stored aseptically at 2°C to 8°C for up to 4 weeks.

Additional information

• A. Reyes, et al. 2015. Xeno-Free and Defined Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells Functionally Integrate in a Large-Eyed Preclinical Model Plaza. Stem Cell Reports.
• Rodin S, et al. 2014. Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment. Nat Commun. 5:3195. doi: 10.1038/ncomms4195.

Material Safety Data Sheet

• LaminStem™ 521 (MSDS)

Manuals and Protocols

• LaminStem™ 521 Protocol
• Weekend-free culture of human pluripotent stem cells on LN-521 (BioLamina)

Product Literature

• Blog Post: Useful tips for culturing hES & hiPS cells with NutriStem® hPSC Medium and LaminStem-521