How can you tell the difference between 3T3 and L1 cells?
Differentiation of 3T3-L1 cells into adipocyte-like cells
- Seed cells in a six-well plate at a density of 3×103 cells per cm2.
- Grow cells in DMEM until a confluency of 70% is reached, changing the medium every 2–3 days.
- To initiate differentiation, remove DMEM and add 2–3 mL MDI induction medium per well (Day 0).
What is 3T3-L1 adipocytes?
Differentiated 3T3-L1 adipocytes are a widely used in vitro model of white adipocytes. In addition to classical white and brown adipocytes that are derived from different cell lineages, beige adipocytes have also been identified, which have characteristics of both white and brown adipocytes.
Are adipocytes terminally differentiated?
Adipogenesis is the formation of adipocytes (fat cells) from stem cells. It involves 2 phases, determination, and terminal differentiation.
How do you culture 3T3-L1 cells?
Place approximately 5,000 cells/cm2 in tissue culture treated cultureware using 3T3-L1 Preadipocyte Medium (PM-1-L1). 5. Maintain cells until they are 100% confluent (in about 6-7 days) in a humidified incubator, 37°C, with 5-10% CO2. Cells will need to be fed every other day with PM-1-L1 during this time.
What is adipogenic differentiation?
Adipogenesis is the process of cell differentiation during which fibroblast-like preadipocytes develop into mature adipocytes. The process is one of the most intensively studied models of cellular differentiation, often using 3T3-L1 preadipocytes as models for study.
What does 3T3-L1 mean?
3T3-L1 is a cell line derived from (mouse) 3T3 cells that is used in biological research on adipose tissue. 3T3-L1 cells have a fibroblast-like morphology, but, under appropriate conditions, the cells differentiate into an adipocyte-like phenotype.
What causes adipocyte differentiation?
Insulin activates its receptor to induce uptake of glucose and free fatty acids by adipocytes. Glucose and free fatty acids are building materials in the biosynthesis of triglycerides, which is stored in the cytoplasm of adipocytes and often used as a marker of adipocyte differentiation.
What does the word adipocyte mean?
Definition of adipocyte : a specialized cell of adipose tissue that stores excess energy in the form of triglyceride droplets and secretes various substances (such as leptin) that play a role in various physiological processes (such as regulation of food intake or lipid metabolism) : fat cell.
How do you thaw 3t3 cells?
Remove the cryovial containing the frozen cells from liquid nitrogen storage and immediately place it into a 37°C water bath. Quickly thaw the cells (< 1 minute) by gently swirling the vial in the 37°C water bath until there is just a small bit of ice left in the vial. Transfer the vial it into a laminar flow hood.
What are Preadipocyte cells?
Preadipocytes are fat cell precursors and are an important cell type in their own right. They comprise a significant proportion of the cells in fat tissue, often outnumbering fat cells, endothelial cells, and macrophages combined.
What are fibroblasts cells?
Fibroblast A fibroblast is a type of cell that contributes to the formation of connective tissue, a fibrous cellular material that supports and connects other tissues or organs in the body. Fibroblasts secrete collagen proteins that help maintain the structural framework of tissues.
What are differentiated 3T3-L1 adipocytes?
Abstract Differentiated 3T3-L1 adipocytes are a widely used in vitromodel of white adipocytes. In addition to classical white and brown adipocytes that are derived from different cell lineages, beige adipocytes have also been identified, which have characteristics of both white and brown adipocytes.
How do you know when adipocytes have been differentiated?
By Day 7–10, fully differentiated adipocyte-like cells should be obtained. Differentiation into adipocyte-like cells can be tracked by Oil Red O staining to monitor lipid accumulation, or by monitoring the expression of adipocyte markers such as adiponectin and FABP4. Protocol adapted from Reed BC, Lane D (1980).
What is 3T3 L1 cell culture used for?
The pre-adipocyte 3T3-L1 cell culture line, derived from mouse embryos, is a widely used model for the study of adipocyte differentiation. Robust and well-characterized, 3T3-L1 pre-adipocytes differentiate and form fat pads indistinguishable from normal adipose tissue when injected into mice.
What is the best way to divide adipocytes?
When splitting cells, wash cells 2x with sterile PBS (-/-) then add 0.05% trypsin and sit at room temperature for 2-5 min. Be careful not to over-trypsinize cells Cells can normally be passaged up to about passage # 25 without problems. Plate cells out in whichever format you require your adipocytes in (i.e. 12 well plates or 15 cm dishes)