Biolaminin 411 LN

Amount	Size	Price
100 µg	1 x 1 ml	€71
1 mg	10 x 1 ml	€625
For larger quantities contact us

Applications Supporting data Instructions Technical info

Recombinant human laminin 511 and laminin 411 can in combination create a natural niche in the culture dish for endocrine pancreatic beta cells, promoting both proliferation of β cells and insulin gene expression.

In the embryo, differentiation and insulin expression by the pancreas β cells is initiated by and dependent on specific signals from capillary endothelial cells (Lammert, 2001). However, pancreatic cells are unable to produce the extracellular proteins laminin 411 and laminin 511, which are expressed and secreted by endothelial cells.

Instead, the pancreatic islets express vascular endothelial growth factor to attract endothelial cells, providing both oxygen and nutrients and form the essential vascular network for insulin production by the β cells (Lammert, 2003; Nikolova, 2006).

Data presented by Nikolova (2006) and colleagues suggest that treating islets with these laminins prior to transplantation will help maintain insulin production until new capillaries are formed in transplanted islets and that laminin 411 and laminin 511 in cell culture potentially could mimic the biological environment in the pancreas.

BioLamina's revolutionary matrix system brings together the unique qualities of laminin 411 and laminin 511 to provide the solution for making pancreatic islet cells thrive.

DIFFERENTIATION AND MAINTENANCE OF PANCREATIC CELLS

One of the main isoform of laminins in capillaries and larger vessels produced by endothelial cells is laminin 411 and, thus, it has a pivotal role in endothelial basal laminae function (Kortesmaa, 2000; Qian, 2007; Wondimu, 2004). Transgenic mice with an α4-chain laminin deficiency exhibit severe disorganization of microvessels, leading to microcirculation abnormalities (Thyboll, 2002; Wang, 2006).

In addition, during sprouting angiogenesis laminins are produced and deposited from the stalk cells to the tip cell filopodia, and control the diameter size of the vessel lumen (Jakobsson, 2008), further supporting the importance of the protein in endothelial structure and function.

Lymphoid cells are derived from bone marrow stem cells and differentiate either in the thymus or in the bone marrow. Lymphocytes are fundamental for innate and adaptive immunity and patrol the body for foreign antigens that activates them. Laminin 411 is secreted by lymphocytes and supports the proliferation, adhesion and migration of lymphocytes (Geberhiwot, 2001).

Laminin 411 is also involved in the survival, adhesion and migration of blood neutrophils (Wondimu, 2004), and works as an adhesive substratum for bone marrow stem and progenitor cells and promote progenitor cell migration in vitro (Gu, 2003).

Cell Culture of Beta Cells

Maintenance and expansion
of insulin producing islets

Endothelial cells

Maturation and maintenance
of blood vessels

Laminins in the pancreas

laminin ECM in pancreatic islets

Islet endothelial cells in mouse pancreas express both laminin 411 (left) and laminin 511 (right) seen as red in these confocal microscopy images. The laminins induce a cell-signaling cascade in the β islet cells, which leads to expression of insulin and cell proliferation.

Laminins by endothelial cells

laminin expression in pancreatic islets

Laminins and other extracellular proteins, including collagen IV, are only produced by islet endothelial cells and not by islet β cells. Laminin 411 and laminin 511 proteins act in concert with instructive and cell-type specific signals and are, in addition, required in liver morphogenesis and growth.

Laminins and insulin

Laminin maintain beta cell insulin expression culture

Laminin 411 and laminin 511 in cell culture can partially substitute the biological environment of the pancreas and can, therefore, potentially help maintain β cell insulin production during islet transplantations before the islets are revascularized by the body.

COATING PROCEDURE

1. Slowly thaw recombinant laminins at +4°C before use. Thawed laminin stock is stable for at least 3 months when stored at +2°C to +8°C under aseptic conditions. For your convenience, the coated plates and diluted coating solution can be kept for up to 4 weeks when stored aseptically at +2°C to +8°C.

2. Dilute the thawed laminin stock solution with 1xDPBS containingCa2+ and Mg2+.

3. Add the diluted laminin solution to tissue culture-treated cultureware for a final coating concentration of 0.5-2 ug/cm2. The optimal coating concentration is cell-dependent. See laminin coating instructions, recommended coating volumes and concentrations.

4. Seal the plate (e.g. with Parafilm®) to prevent evaporation and incubate at +2°C to +8°C overnight. If a more rapid coating is required, incubate at +37°C for 2 hours. Make sure the laminin solution is spreaded evenly across the surface. Note that the laminin matrix will be inactivated if let dry.

IMPORTANT NOTES

The Laminin stock solution is stable for 3 years when stored at -20°C to -80°C. Repeated freeze thawing should be avoided. If desired, the laminin stock can be dispensed into working aliquots and stored at -20°C to -80°C.
The protocol can easily be made totally defined and xeno-free with your choice of culture medium and enzyme.
Before start, all solutions used for cell passaging should be aliquoted in sufficient amounts and pre-warmed at +37°C, 5% CO2.
Coat the wells in advance with the laminin solution according to the instruction here.

Product name

Biolaminin 411 LN

Product code

LN411

Declaration

For research use only

Documents

MSDS

CoA

Storage

-20°C to -80°C

Stock concentration

0.1mg/ml

Appearance

Clear, colourless, buffered solution with a
pH of 7.2 with 10% glycerol and 0.02% NaN3

Shipping Condition

Dry Ice

Protein name

Laminin 411 (Laminin-8)

Classification

Defined and xeno-free, human
recombinant protein

Product application

Culture of pancreatic islet cells, endothelial cells, adipocytes and epidermal cells