Primary cells of human cerebellar granule

Primary cells of human cerebellar granule

Training Information:

Package conditionsPLL（0.1mg/ml）

culture mediumcontainFBSGrowth additivesPenicillinTheStreptomycinwait

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

Growth characteristics: Wall adhesion

Cell morphology, neuronal cell like

Passage characteristics belong to terminally differentiated cells; Belonging to non proliferating cell population

Digestive fluid0.25%

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

Human cerebellar granule cells were isolated from the cortical tissue of the small brain; Neurons are the basic structural and functional units of the nervous system, and cerebellar granule neurons are smaller neurons with a diameter of approximately10MmIt is very abundant in the central nervous system, accounting for almost half of the number of neurons. Due to the similarity in growth and differentiation between cerebellar neurons and cortical neurons, as well as their large quantity and ease of sampling, cerebellar granule neurons are important tools for studying neuronal growth and development, axonal regeneration, and the mechanisms of neurological disease occurrence, as well as clinical neuropharmacology. The cerebellar granule neurons are the main interneurons in the cerebellum, and are abundant in the cerebellum of mammals. The axons of granule neurons are connected to mossy fibers and crawling fibers, forming neuronal circuits in the cerebellar cortex, which play a very important role in the neural activity of the cerebellum. In animal transmissible spongiform encephalopathy, lesions can affect cerebellar granule neurons, causing damage to neuronal circuits in the cerebellar cortex, resulting in neurological behavioral disorders. The study of the response of cerebellar granule neurons in the pathological changes of animal infectious spongiform encephalopathy, the mechanism of pathological occurrence, especially the molecular mechanism, relies on the establishment of cerebellar granule neuron cell models. The axons of cerebellar granule cells are parallel fibers distributed along the coronal axis, which ensures the unidirectional conduction of excitation. This is a key hypothesis in cerebellar functional theory, and cerebellar granule cells pass throughg-Aminobutyric acid receives inhibitory synaptic information transmission from Golgi cells.

Method Introduction:

The human cerebellar granule cells isolated in the company's laboratory were prepared by protease digestion combined with neuron specific culture medium and chemical reagent inhibition screening. The total number of cells is approximately5×10?cells/Bottle.

Quality inspection:

The human cerebellar granule cells isolated from the company laboratory were subjected to β - κ B treatment-TubulinImmunofluorescence identification, purity can reach90%Above, and not containingHIV-1TheHBVTheHCVMycoplasma, bacteria, yeast, fungi, etc.

① Organizational block cultivation method

Tissue block culture is a commonly used, simple, and highly successful primary culture method. The basic method is to inoculate the cut small tissue clusters into a culture bottle (or dish), and the bottle wall can be pre coated with a thin layer of collagen to facilitate the adhesion of the tissue blocks to the bottle wall, allowing surrounding cells to grow outward along the bottle wall.

② Digestive cultivation method

③ Suspension cell culture method

For cells that grow in suspension, such as leukemia cells, lymphocytes, bone marrow cells, cancer cells and immune cells in pleural and ascites, digestion is not required. They can be isolated by low-speed centrifugation and cultured directly, or cultured by inoculating after lymphocyte stratification.

④ Organ culture

Organ culture refers to the direct cultivation of organs or tissue blocks obtained from donors under specific environmental conditions outside the body without tissue separation. Organ culture can maintain the relative integrity of organ tissues and can be used to focus on observing the connections, arrangements, and interactions between cells, as well as the biological regulatory effects of local environments.

After receiving the cells, please proceed with the following steps for the customer. (The number of passages for this cell is limited to about 2. It is recommended that the customer arrange subsequent experiments as soon as they receive the cells.)

1. Remove the cell vial, disinfect it with 75% alcohol, remove the sealing film, and place it in a 37 ℃, 5% CO2 incubator for 6-8 hours to stabilize the cell state.

2. When the cells reach 80% confluence, prepare for subculture.

3. Cell passage:

1) Suck out the culture medium from the cell vial and wash the cells once with PBS.

2) Add about 1mL of 0.125% digestive solution to the culture bottle and digest at 37 ℃ for about 3 minutes; Under the microscope, after the cells shrink and become round, discard the digestion solution and add culture medium to terminate digestion.

3) Gently blow and mix with a straw, inoculate and passage in appropriate ratios such as 1:2 or 1:3, then add fresh culture medium to 5mL and culture in a 37 ℃ 5% CO2 cell incubator.

4) After the cells adhere to the wall, culture and observe, and replace with fresh culture medium every 2-3 days.

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