neuronal enrichment of dorsal root ganglia by immunopanning

Neuronal Enrichment of Dorsal Root Ganglia by Immunopanning

For cell layering, add 2 milliliters of 15% BSA, and gently coat the sides of the closed tube. To remove the myelin debris, carefully layer the cell suspension 1 milliliter at a time by pipetting against the side of the tube, on top of the BSA cushion.
Centrifuge the tube at 300 g for 10 minutes at room temperature, with slow acceleration and deceleration. The middle myelin layer shows flakes of white material. Using a P1000 pipette, remove the clear liquid on top, the myelin phase in between, and the BSA, 1 milliliter at a time, leaving approximately 100 microliters, so as not to disturb the pellet.
For antigen retrieval, add 5 milliliters of panning buffer and incubate the tube in a 37 degrees Celsius and 10% carbon dioxide incubator for 30 to 45 minutes. Next, wash the incubated CD45 dish three times with DPBS, and pour off the final rinse. Then, pour the cell suspension into the CD45 dish.
Incubate the dish for 20 minutes at room temperature, swirling gently at 10-minute intervals, to allow the cells to gain equal access to the antibody. Gently shake the CD45 dish, and prop it up at an angle. Carefully pipette 1 milliliter of the cell suspension over the dish to collect unbound cells. Then, transfer it to the PDGFR-beta dish, previously washed three times with DPBS, and incubate.
Transfer the unbound cells from the PDGFR-beta dish to the O4 dish, as demonstrated earlier, and incubate the plate. After the incubation, transfer the cell suspension to a 15-milliliter conical tube. Remove the laminin, and immediately, plate the cells at the desired density without allowing them to dry before incubating the plate.

neuronal-enrichment-of-dorsal-root-ganglia-by-immunopanning

Nanyang Technological University

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1. Experimental method
<ol>
	<li>Day 1: preparation of dishes
	<ol>
		<li>In a biosafety cabinet, coat the desired culture surface with enough poly-D-lysine to cover the bottom of the well. Store overnight at 4 &#176;C. Here, 24-well glass-bottom plates were used for imaging, and thus 300 &#181;L of poly-D-lysine was used.</li>
		<li>In a biosafety cabinet, prepare the panning dishes: for the CD45 dish, add 10 mL of working tris-HCl solution and 30 &#181;L of goat anti-rat IgG (immunoglobulin G) to a 10 cm Petri dish; for the PDGFR&#946; (platelet-derived growth factor receptor beta) dish, add 10 mL of working tris-HCl solution and 30 &#181;L of goat anti-rabbit IgG to a 10 cm Petri dish; for the O4 hybridoma dish, add 10 mL of working tris-HCl solution and 30 &#181;L of goat anti-mouse IgG to a 10 cm Petri dish. Ensure the solution covers the entire dish area and leave it at 4 &#176;C overnight.</li>
	</ol>
	</li>
	<li>Day 2: dissection, trituration, and immunopanning
	<ol>
		<li>Aspirate the poly-D-lysine, wash the plates three times with tissue culture water, tilt the lid open, and allow the surface to dry completely in a biosafety cabinet. Apply enough laminin to the plates to coat the bottom of each well, and place in an incubator at 37 &#176;C. For the 24-well plates, 200 &#181;L is sufficient.</li>
		<li>Finish preparing the panning dishes. Wash each dish with D-PBS and incubate with 10 &#181;g of primary antibody. Allow to sit at room temperature for at least 2 h.</li>
		<li>For the CD45 dish, add 5 mL of 0.2% BSA (bovine serum albumin) and 20 &#181;L of CD45 primary; for the PDGFR&#946; dish, add 5 mL of 0.2% BSA and 15.4 &#181;L of PDGFR&#946;; for the O4 dish, add 3 mL of 0.2% BSA, 2 mL of O4 hybridoma, and an additional 100 &#181;L of 4% BSA (to ensure the final BSA concentration remains at 0.2%).</li>
		<li>Euthanize one to four mice using 0.1 mL/kg sodium pentobarbitol (per 10 cm immunopanning dish). We used C57BL/6 mice at 8-10 weeks old. Both sexes were used and cultured separately. Perfuse the animal with 30 mL of cold 0.9% saline.</li>
		<li>Pin the front and hind paws to a styrofoam stage and use a clean razor blade to expose the spinal column from the back. Perform a laminectomy by making cuts about halfway deep on either side of the dorsal spinal column, removing the dorsal half of the column to expose the spinal cord. Either gently cut the nerves and remove the spinal cord or gently push the cord to the side, maintaining the nerve integrity.</li>
		<li>Use the spinal nerve roots (either severed or attached to the spinal cord) to find and remove all the DRGs (dorsal root ganglions) from either side of the spinal column. Trim the nerve debris from each DRG as it is removed (more nerve debris in the culture may lead to poor culture survival). Collect the DRGs in 15 mL of HBSS (Hank&#8217;s balanced salt solution in a 15 mL conical tube on ice. 
		NOTE: Note that the tissue is dissected in open air, but once the DRGs are added to the tube, they should be treated as sterile and only manipulated in a biosafety cabinet.</li>
		<li>Place frozen stocks of stemxyme 1 and 0.4% DNAse in a water bath approximately 30 min prior to the end of the dissections (roughly when starting the last mouse). Allow them to settle to the bottom of the conical tube, then continue the protocol in a biosafety cabinet.</li>
		<li>Aspirate most of the HBSS-/- from the DRGs. Use a pipette rather than suction to get &#60;1 mL of liquid, and leave ~100 &#181;L liquid so as not to risk losing tissue.</li>
		<li>Wash three times with 1 mL of HBSS-/-. Add 5 mL of working stemxyme solution to the tissue. Cover the lid with a transparent film and float the tube on its side in a water bath set to 37 &#176;C for 1 h.</li>
		<li>After incubation in the enzyme, add 1 mL of low ovo inhibitor solution to the tube. Triturate the cells gently with a p1000 pipette 10-15 times. Allow the tissue chunks to settle.</li>
		<li>Transfer the top 2-3 mL of solution (containing dissociated cells) to fresh low ovomucoid solution. Repeat until the tissue is fully dissociated and no visible chunks remain.</li>
		<li>Centrifuge for 10 min at 300 x g at room temperature. Siphon off the supernatant again, using a pipette rather than suction for the last 1 mL, leaving ~100 &#181;L, and resuspend the cell pellet in 1 mL of panning buffer.</li>
		<li>Gently pipette to mix. Pre-wet a 70 &#181;m cell strainer with 1 mL of panning buffer over a 50 mL conical tube. Filter the cell solution through the cell strainer.</li>
		<li>Wash the tube with 1 mL of panning buffer, then pass through the strainer (3 mL of filtrate). Layer the cell suspension gently (1 mL at a time) on top of 2 mL of a 15% BSA cushion to remove myelin debris. A 2 mL cushion is effective for two mice, and 3 mL is effective for four mice.
		<ol>
			<li>For layering, place the 2 mL of BSA in the tube, close it, and gently coat the sides of the tube with BSA. Then, gently layer the cell suspension on top by pipetting it against the side of the tube.</li>
		</ol>
		</li>
		<li>Centrifuge at room temperature at 300 x g for 10 min (slow acceleration and deceleration). Use a P1000 pipette to remove the clear liquid on top, the myelin phase in between, and the BSA, 1 mL at a time (changing tips in between each mL). Leave ~100 &#181;L of BSA so as not to disturb the pellet.</li>
		<li>Add 5 mL of panning buffer and incubate the tube in a 37 &#176;C, 10% CO2 incubator for 30-45 min. This allows for antigen retrieval. Be sure to loosen the cap to allow gas exchange.</li>
		<li>Wash the CD45 dish three times with D-PBS. Pour off the final rinse. Decant the cell suspension into the CD45 dish.</li>
		<li>Incubate the cells on the CD45 dish for a total of 20 min at room temperature, swirling gently at the 10 min point to allow the cells equal access to the antibody.</li>
		<li>Rinse the PDGFR&#946; dish three times with D-PBS and pour off the final rinse. Transfer the unbound cells from the CD45 dish to the PDGFR&#946; dish.</li>
		<li>Gently shake the CD45 dish and prop it up at an angle. Gently pipette 1 mL of the cell suspension over the dish to collect unbound cells. Decant it into the PDGFR&#946; dish and pipette to transfer the remaining cell suspension to the PDGFR&#946; plate.</li>
		<li>Incubate the PDGFR&#946; plate for a total of 20 min at room temperature, swirling gently at the 10 min point to allow the cells equal access to the antibody.</li>
		<li>Rinse the O4 dish three times with D-PBS and pour off the final rinse. Transfer the unbound cells from the PDGFR&#946; dish to the O4 dish using the same method as in step 1.2.19. Incubate the O4 plate for a total of 20 min at room temperature, swirling gently at the 10 min point to allow the cells equal access to the antibody.</li>
		<li>Transfer the cell suspension to a 15 mL conical tube and centrifuge for 10 min at 300 x g. Resuspend the cells in the desired volume and dilute 1:1 with trypan blue for cell viability. Count medium-to-large cells with a hemocytometer (this will allow for the best representation of the number of neurons in the culture).</li>
		<li>Remove the laminin and plate the cells at the desired density. Do not allow the plate to dry between laminin removal and plating, and add the cells immediately.</li>
		<li>Culture the quantified cells for neuronal enrichment (Figure 1) with 500 neurons in 25 &#181;L of neural medium (as described below) per well in 24-well plates and incubate at 37 &#176;C for 30 min. Flood the wells to a final volume of 500 &#181;L.
		<ol>
			<li>Prepare 50 mL of neuron media by adding 23.2 mL of DMEM, 23.2 mL of neurobasal medium, 500 &#181;L of glutamax, 500 &#181;L of sodium pyruvate, 500 &#181;L of penicillin/streptomycin (aliquot immediately upon arrival and store at -20 &#176;C), 500 &#181;L of insulin (5 &#181;g/mL final), 500 &#181;L of SATO, 50 &#181;L of N-acetyl-L-cysteine (NAC; 5 &#181;g/mL final), and 1,000 &#181;L of B27+ (immediately aliquot and store at -20 &#176;C upon receiving).
			<ol>
				<li>Prepare the insulin stock by dissolving it in culture-grade water to a concentration of 0.5 mg/mL (50 mg in 100 mL), adjusting the pH to 7.4, filter sterilizing (0.22 &#181;m), preparing 500 &#181;L aliquots, and storing at -20 &#176;C.</li>
				<li>Prepare the SATO stock by combining 20 &#181;L of progesterone (2.5 mg in 100 &#181;L of ethanol), 800 &#181;L of sodium selenite (4 mg in 10 mL of neurobasal media + 10 &#181;L of 1 N NaOH (Sodium hydroxide)), 800 mg of BSA, 800 mg of transferrin, 128 mg of putrescine dihydrochloride, and 80 mL of neurobasal. Filter sterilize (0.22 &#181;m), prepare 500 &#181;L aliquots, and store at -20 &#176;C. It is essential to make fresh progesterone and sodium selenite solutions.</li>
				<li>Prepare 50 &#181;L of N-acetyl-L-cysteine (NAC) stock by dissolving in neurobasal medium to a stock concentration of 5 mg/mL (50 mg in 10 mL), filter sterilize (0.22 &#181;m), aliquot, and store at -20 &#176;C.</li>
			</ol>
			</li>
		</ol>
		</li>
	</ol>
	</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.2 um Syringe filters</td>
			<td>Fisher</td>
			<td>723-2520</td>
			<td></td>
		</tr>
		<tr>
			<td>100 mm petri dishes</td>
			<td>ThermoFisher</td>
			<td>FB0875712</td>
			<td></td>
		</tr>
		<tr>
			<td>24 well black glass-bottom plates</td>
			<td>Cellvis</td>
			<td>P24-1.5H-N</td>
			<td></td>
		</tr>
		<tr>
			<td>70 um cell strainer</td>
			<td>Cedarlane</td>
			<td>15-1070-1(BI)</td>
			<td></td>
		</tr>
		<tr>
			<td>B27+ supplement</td>
			<td>Gibco</td>
			<td>A3582801</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin</td>
			<td>Sigma Aldrich</td>
			<td>A7906</td>
			<td>for 15% BSA cushion, and ICC (heat shock fraction &#8805;98%)</td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin</td>
			<td>Sigma Aldrich</td>
			<td>A4161</td>
			<td>for 4% BSA for immunopanning, and SATO for media (essentially globulin free, suitable for cell culture, &#8805;99%)</td>
		</tr>
		<tr>
			<td>CD45 antibody</td>
			<td>BD Pharmigen</td>
			<td>550539</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Invitrogen</td>
			<td>D1306</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM</td>
			<td>Gibco</td>
			<td>11960069</td>
			<td></td>
		</tr>
		<tr>
			<td>DNAse</td>
			<td>Worthington</td>
			<td>LS002007</td>
			<td>for stemxyme solution and panning buffer</td>
		</tr>
		<tr>
			<td>D-PBS</td>
			<td>Sigma Aldrich</td>
			<td>D8662</td>
			<td></td>
		</tr>
		<tr>
			<td>EBSS</td>
			<td>Sigma Aldrich</td>
			<td>E6267</td>
			<td>for DNAse solution</td>
		</tr>
		<tr>
			<td>Filter paper P8 grade</td>
			<td>ThermoFisher</td>
			<td>09-795K</td>
			<td>for 8% PFA</td>
		</tr>
		<tr>
			<td>Glutamax</td>
			<td>ThermoFisher</td>
			<td>35050061</td>
			<td></td>
		</tr>
		<tr>
			<td>goat-anti-mouse IgG</td>
			<td>Jackson Immunoresearch</td>
			<td>115-005-020</td>
			<td>for O4 dish</td>
		</tr>
		<tr>
			<td>goat-anti-rabbit 488</td>
			<td>Invitrogen</td>
			<td>A11008</td>
			<td></td>
		</tr>
		<tr>
			<td>goat-anti-rabbit IgG</td>
			<td>Jackson Immunoresearch</td>
			<td>115-005-003</td>
			<td>for PDGFRB dish</td>
		</tr>
		<tr>
			<td>goat-anti-rat IgG</td>
			<td>Jackson Immunoresearch</td>
			<td>115-005-044</td>
			<td>for CD45 dish</td>
		</tr>
		<tr>
			<td>HBSS -/-</td>
			<td>Sigma Aldrich</td>
			<td>14175145</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulin</td>
			<td>Sigma Aldrich</td>
			<td>I2643</td>
			<td></td>
		</tr>
		<tr>
			<td>laminin</td>
			<td>Invitrogen</td>
			<td>23017-015</td>
			<td></td>
		</tr>
		<tr>
			<td>N-acetyl cysteine</td>
			<td>Sigma Aldrich</td>
			<td>A8199</td>
			<td></td>
		</tr>
		<tr>
			<td>Neurobasal</td>
			<td>Gibco</td>
			<td>21103049</td>
			<td></td>
		</tr>
		<tr>
			<td>Normal Donkey Serum</td>
			<td>Sigma-Aldrich</td>
			<td>D9663</td>
			<td></td>
		</tr>
		<tr>
			<td>O4 antibody</td>
			<td>n/a</td>
			<td>n/a</td>
			<td>Hybridoma</td>
		</tr>
		<tr>
			<td>Ovomucoid trypsin inhibitor</td>
			<td>Cedarlane</td>
			<td>LS003086</td>
			<td>for low ovo</td>
		</tr>
		<tr>
			<td>paraformaldehyde prills</td>
			<td>Sigma Aldrich</td>
			<td>441244</td>
			<td>for 8% PFA</td>
		</tr>
		<tr>
			<td>PDGFRB antibody</td>
			<td>Abcam</td>
			<td>AB32570</td>
			<td></td>
		</tr>
		<tr>
			<td>penicillin/ streptomycin</td>
			<td>Gibco</td>
			<td>15140-122</td>
			<td></td>
		</tr>
		<tr>
			<td>Poly-D-Lysine</td>
			<td>Sigma Aldrich</td>
			<td>P6407</td>
			<td></td>
		</tr>
		<tr>
			<td>Progesterone</td>
			<td>Sigma Aldrich</td>
			<td>P8783</td>
			<td>for SATO</td>
		</tr>
		<tr>
			<td>Putrescine dihydrochrloride</td>
			<td>Sigma Aldrich</td>
			<td>P5780</td>
			<td>for SATO</td>
		</tr>
		<tr>
			<td>Sodium phosphate dibasic</td>
			<td>Fisher</td>
			<td>S374-1</td>
			<td>for 0.2 M PB</td>
		</tr>
		<tr>
			<td>Sodium Phosphate monobasic dihydrate</td>
			<td>Sigma Aldrich</td>
			<td>4269</td>
			<td>for 0.2 M PB</td>
		</tr>
		<tr>
			<td>Sodium Pyruvate</td>
			<td>ThermoFisher</td>
			<td>11360070</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Selenite</td>
			<td>Sigma Aldrich</td>
			<td>S5261</td>
			<td>for SATO</td>
		</tr>
		<tr>
			<td>Stemxyme I</td>
			<td>Cedarlane</td>
			<td>LS004107</td>
			<td>for tissue dissociation; combination collagenase</td>
		</tr>
		<tr>
			<td>Transferrin</td>
			<td>Sigma Aldrich</td>
			<td>T1147</td>
			<td>for SATO</td>
		</tr>
		<tr>
			<td>Tris-HCl</td>
			<td>Millipore Sigma</td>
			<td>T5941</td>
			<td></td>
		</tr>
		<tr>
			<td>trypan blue</td>
			<td>Gibco</td>
			<td>15250-061</td>
			<td></td>
		</tr>
		<tr>
			<td>&#946;3-Tubulin</td>
			<td>Sigma-Aldrich</td>
			<td>T2200</td>
			<td></td>
		</tr>
	</tbody>
</table>

Source: Maguire, A. D., et al. <a href="https://app-jove-com.remotexs.ntu.edu.sg/v/64603">Enrichment of Adult Mouse Dorsal Root Ganglia Neuron Cultures by Immunopanning.</a> J. Vis. Exp. (2023)
This video demonstrates a protocol for enriching neuronal cultures from dorsal root ganglia by immunopanning, thereby improving their purity and utility for neurological research.

<img alt="Figure 1" src="/files/ftp_upload/22584/22584_Figure 1.jpg" /> 
Figure 1: DRG culture neuronal enrichment by immunopanning. Fixed cultures were stained with &#946;3-tubulin for neurons and DAPI for all nuclei. (A) Representative image of a whole (non-immunopanned) culture. (B) Representative image of an immunopanned culture. (C) Quantification of &#946;3-tubulin-positive cells as a percentage of total DAPI-positive cells. Bars indicate mean &#177; standard error mean (SEM). **** = p &#60; 0.0001, unpaired t-test.

1. Experimental method

	Day 1: preparation of dishes
	
		In a biosafety cabinet, coat the desired culture surface with enough poly-D-lysine to cover the ...

Begin by layering the dorsal root ganglia-derived cells onto bovine serum albumin or BSA.
The cell suspension contains neurons, immune cells, fibroblasts, Schwann cells, and fatty myelin debris.
Centrifuge. The denser DRG cells pass through, forming a pellet.
Remove the myelin layer along with the serum.
Resuspend the cells in an immunopanning buffer, which improves the accessibility of the antigen for antibodies.
Take Petri dishes coated with secondary antibodies conjugated with cell-specific primary antibodies.
Add the cell suspension to the immune cell-specific antibody-coated Petri dish.
The immune cells attach to the antibodies while the other cells remain unbound.
Transfer the remaining cells to the next Petri dish to remove the fibroblasts.
Repeat the procedure to remove the Schwann cells.
Plate the unbound cells onto a matrix-coated multiwell plate containing a neurobasal medium and incubate to obtain an enriched neuronal culture.

16 次观看. 通过免疫淘选实现背根神经节的神经元富集

视频: 通过免疫淘选实现背根神经节的神经元富集 - 实验

Dorsal Root Ganglia Isolation and Primary Culture to Study Neurotransmitter Release

Dorsal root ganglia (DRG) contain cell bodies of sensory neurons. This type of neuron is pseudo-unipolar, with two axons that innervate peripheral tissues, such as skin, muscle and visceral organs, as well as the spinal dorsal horn of the central nervous system. Sensory neurons transmit somatic sensation, including touch, pain, thermal, and proprioceptive sensations. Therefore, DRG primary cultures are widely used to study the cellular mechanisms of nociception, physiological functions of sensory neurons, and neural development. The cultured neurons can be applied in studies involving electrophysiology, signal transduction, neurotransmitter release, or calcium imaging. With DRG primary cultures, scientists may culture dissociated DRG neurons to monitor biochemical changes in single or multiple cells, overcoming many of the limitations associated with in vivo experiments. Compared to commercially available DRG-hybridoma cell lines or immortalized DRG neuronal cell lines, the composition and properties of the primary cells are much more similar to sensory neurons in tissue. However, due to the limited number of cultured DRG primary cells that can be isolated from a single animal, it is difficult to perform high-throughput screens for drug targeting studies. In the current article, procedures for DRG collection and culture are described. In addition, we demonstrate the treatment of cultured DRG cells with an agonist of neuropeptide FF receptor type 2 (NPFFR2) to induce the release of peptide neurotransmitters (calcitonin gene-related peptide (CRGP) and substance P (SP)).

Dorsal root ganglia (DRG) primary cultures are frequently used to study physiological functions or pathology-related events in sensory neurons. Here, we demonstrate the use of lumbar DRG cultures to detect the release of neurotransmitters after neuropeptide FF receptor type 2 stimulation with a selective agonist.

背根神经节分离与原代培养对神经递质释放的研究

Dorsal root ganglia (DRG) contain cell bodies of sensory neurons. This type of neuron is pseudo-unipolar, with two axons that innervate peripheral ...

科研

JoVE Journal

神经科学

Purification of Fibroblasts and Schwann Cells from Sensory and Motor Nerves in Vitro

The principal cells in the peripheral nervous system are the Schwann cells (SCs) and the fibroblasts. Both these cells distinctly express the sensory and motor phenotypes involved in different patterns of neurotrophic factor gene expression and other biological processes, affecting nerve regeneration. The present study has established a protocol to obtain highly purified rat sensory and motor SCs and fibroblasts more rapidly. The ventral root (motor nerve) and the dorsal root (sensory nerve) of neonatal rats (7-days-old) were dissociated and the cells were cultured for 4-5 days, followed by isolation of sensory and motor fibroblasts and SCs by combining differential digestion and differential adherence methods sequentially. The results of immunocytochemistry and flow cytometry analyses showed that the purity of the sensory and motor SCs and fibroblasts were &#62;90%. This protocol can be used to obtain a large number of sensory and motor fibroblasts/SCs more rapidly, contributing to the exploration of sensory and motor nerve regeneration.

Here, we present a method to purify fibroblasts and Schwann cells from sensory and motor nerves in vitro.

从体外感觉和运动神经中纯化成纤维细胞和施万细胞

The principal cells in the peripheral nervous system are the Schwann cells (SCs) and the fibroblasts. Both these cells distinctly express the sensory and ...

In Vitro Myelination of Peripheral Axons in a Coculture of Rat Dorsal Root Ganglion Explants and Schwann Cells

The process of myelination is essential to enable rapid and sufficient signal transduction in the nervous system. In the peripheral nervous system, neurons and Schwann cells engage in a complex interaction to control the myelination of axons. Disturbances of this interaction and breakdown of the myelin sheath are hallmarks of inflammatory neuropathies and occur secondarily in neurodegenerative disorders. Here, we present a coculture model of dorsal root ganglion explants and Schwann cells, which develops a robust myelination of peripheral axons to investigate the process of myelination in the peripheral nervous system, study axon-Schwann cell interactions, and evaluate the potential effects of therapeutic agents on each cell type separately. Methodologically, dorsal root ganglions of embryonic rats (E13.5) were harvested, dissociated from their surrounding tissue, and cultured as whole explants for 3 days. Schwann cells were isolated from 3-week-old adult rats, and sciatic nerves were enzymatically digested. The resulting Schwann cells were purified by magnetic-activated cell sorting and cultured under neuregulin and forskolin-enriched conditions. After 3 days of dorsal root ganglion explant culture, 30,000 Schwann cells were added to one dorsal root ganglion explant in a medium containing ascorbic acid. The first signs of myelination were detected on day 10 of coculture, through scattered signals for myelin basic protein in immunocytochemical staining. From day 14 onward, myelin sheaths were formed and propagated along the axons. Myelination can be quantified by myelin basic protein staining as a ratio of the myelination area and axon area, to account for the differences in axonal density. This model provides experimental opportunities to study various aspects of peripheral myelination in vitro, which is crucial for understanding the pathology of and possible treatment opportunities for demyelination and neurodegeneration in inflammatory and neurodegenerative diseases of the peripheral nervous system.

In the coculture system of dorsal root ganglia and Schwann cells, myelination of the peripheral nervous system can be studied. This model provides experimental opportunities to observe and quantify peripheral myelination and to study the effects of compounds of interest on the myelin sheath.

大鼠背根神经节外植体和雪旺细胞共培养中外周轴突的体外髓鞘形成

The process of myelination is essential to enable rapid and sufficient signal transduction in the nervous system. In the peripheral nervous system, ...

Neuronal Enrichment of Dorsal Root Ganglia by Immunopanning

成績單

Neuronal Enrichment of Dorsal Root Ganglia by Immunopanning