Chicken skeletal muscle cells

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Product Name:Chicken skeletal muscle cells

Organizational source:Skeletal muscle tissue

Product Specifications5×105cells/T25cell culture flask

Training Information:

Package conditionsPLL（0.1mg/ml）

The culture medium containsFBSGrowth additivesPenicillinTheStreptomycinwait

Fluid change frequency per2-3Change the fluid once every day

Growth characteristics: Wall adhesion

Cell morphology fibroblast like

Passage characteristics can be passed down2-3generation

digestive juice0.25%

Cultivation conditions gas phase: air,95%；CO2，5%

Cell Introduction:

Chicken skeletal muscle isolated from limb muscle tissue; Skeletal muscle, also known as striated muscle, is a type of muscle that accounts for approximately one percent of the body weight40%Skeletal muscle fibers are long cylindrical multinucleated cells, with a basement membrane tightly attached to the outer surface of the muscle membrane. Belonging to striated muscles, striated muscles also include myocardial and visceral striated muscles, among which skeletal muscles are mainly distributed in the limbs. Each muscle is an organ with a certain shape, structure, and function, with abundant blood vessels and lymphatic distribution. It contracts or relaxes under the control of somatic nerves and performs voluntary movements. Muscles can be named based on their common shape, size, position, starting and ending points, fiber direction, and function. Named according to their morphology, such as trapezius muscle, rhombus muscle, deltoid muscle, piriformis muscle, etc. Skeletal muscle cells are fibrous, non branching, with obvious horizontal stripes, and many nuclei located below the cell membrane. There are many filamentous myofibrils arranged parallel to the long axis of muscle cells. Each myofibril has alternating bright bands (I-bands) and dark bands（AWith). Bright band staining is lighter, while dark band staining is darker. There is a brighter line in the middle of the dark band calledHLine.HThere is one in the middle of the lineMLine. In the middle of the bright band, there is a darker line calledZLine. twoZThe section between the lines is called a sarcomere. Skeletal muscle cells, also known as striated muscle cells, are fibrous, non branching, with obvious striations, many nuclei, and are located below the cell membrane. The cells are mostly spindle shaped and have a certain degree of directionality. Primary isolation and culture of skeletal muscle cells3After the day, cells can be seen to adhere to the wall and extend, with varying shapes and sizes of cells, including spindle shaped, irregular shaped, triangular or fan-shaped, and oval shaped and centered nuclei;2After the Zhou period, the cells converge and most of them extend in a long spindle shape with abundant cytoplasm and branching protrusions. The cells are arranged in parallel as a single layer or partially overlapping multiple layers, with ups and downs; When the cell density is low, it often interweaves into a network; When the density is high, it is arranged in a vortex or fence shape. After passage, the cells grow rapidly,4-6The sky can converge and maintain the above morphological and growth characteristics.

Method Introduction:

The chicken skeletal muscle isolated in the company's laboratory was prepared using a mixed collagenase digestion method combined with differential adhesion method, with a total cell count of approximately5×10?cells/Bottle.

Quality inspection:

The chicken skeletal muscle isolated in the company's laboratory was subjected to alpha irradiation-Sarcometric actinImmunofluorescence identification, purity can reach90%Above, and not containingHIV-1TheHBVTheHCVMycoplasma, bacteria, yeast, fungi, etc.

Cultivation steps:

1、 Resuscitate cells: Quickly shake and thaw a cryovial containing 1mL of cell suspension in a 37 ℃ water bath, then add 5mL of culture medium and mix well. Centrifuge at 1000 RPM for 5 minutes, discard the supernatant, add 4-6 mL of culture medium, and blow evenly. Then add all cell suspensions to the culture bottle and culture overnight (or add the cell suspensions to a 6cm dish), and culture overnight. The next day, change the solution and check the cell density.

2、 Cell passage: If the cell density reaches 80% -90%, passage culture can be carried out.

a）、 For adherent cells, the following methods can be used for passaging:

1. Discard the culture supernatant and rinse the cells 1-2 times with PBS that does not contain calcium or magnesium ions.

2. Add 1-2ml of digestion solution (0.25% Trypsin 0.53mM EDTA) to a culture bottle, place it in a 37 ℃ incubator for digestion for 1-2 minutes, and then observe the cell digestion under a microscope. If most of the cells become round and fall off, quickly take them back to the operating table, lightly tap the culture bottle a few times, and add 5ml or more of culture medium containing 10% serum to terminate digestion.

3. Gently blow the cells, remove and aspirate, centrifuge at 1000RPM for 8-10 minutes, discard the supernatant, add 1-2mL of culture medium, and blow evenly.

4. Add 5-6ml/bottle of culture medium and divide the cell suspension into new dishes or bottles containing 5-6 ml of culture medium in a ratio of 1:2.

b）、 For suspended cells, the following methods can be used for passaging:

Method 1: Collect cells, centrifuge at 1000RPM for 8-10 minutes, discard the supernatant, add 1-2ml of culture medium and blow evenly. Divide the cell suspension into new dishes or bottles containing 8ml of culture medium in a ratio of 1:2 to 1:5.

Method 2: You can choose the method of half changing the medium. After discarding half of the medium, suspend the remaining cells and divide the cell suspension into new dishes or bottles containing 8ml of medium in a ratio of 1:2 to 1:3.
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How to handle received cells?

1. Firstly, observe whether the cell culture bottle is intact and whether there is any leakage or turbidity in the culture medium. If there are any, please take photos and contact technical support in a timely manner (the photos taken will be used as the basis for subsequent services).

2. Wipe the surface of the cell culture bottle with 75% alcohol and observe the cell status under a microscope. Due to transportation issues, a small amount of adherent cells may detach from the bottle wall; Do not open the culture bottle cap yet, place the cells in the cell culture box and let them stand for 2-4 hours to stabilize the cell state.

3. Carefully read the cell manual to understand cell related information, such as adhesion characteristics (adhesion/suspension), cell morphology, basic culture medium used, serum ratio, required cytokines, passage ratio, fluid exchange frequency, etc.

4. After settling, take out the cell culture bottle, examine it under a microscope, take photos, and record the cell status (the photos taken will be used as the basis for subsequent services); It is recommended to regularly take photos and record the growth status of cells after subculture.

5. Adherent cells: If the cell growth density exceeds 80%, it can be passaged normally; If the cell density does not exceed 80%, remove the culture medium from the cell culture bottle and reserve about 5ml for further cultivation until the cell density reaches around 80% before proceeding with the passage operation. The bottle cap can be slightly loosened.

6. Suspended cells: Transfer all the liquid in the cell culture bottle to a 50ml sterile centrifuge tube and centrifuge at 1200rpm for 5 minutes. After centrifugation, collect the supernatant culture medium for later use. Add 5ml of culture medium to the bottom of the tube and resuspend the cell precipitate. During microscopic examination, if the cell density exceeds 80%, the cell suspension can be cultured in two cell culture bottles and supplemented with culture medium up to 5ml. If the cell density does not exceed 80%, the cell suspension can be transferred to the original bottle for further cultivation until the cell density reaches around 80% before passaging