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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Graber, David J. Harris, Brent T. |
| Description | Country affiliation: United States Author Affiliation: Graber DJ ( Dartmouth Medical School, Department of Pathology, Lebanon, NH 03756, USA.) |
| Abstract | We describe an immunopanning protocol to isolate, enrich, and culture spinal motor neurons from rat embryonic spinal cords. The method takes advantage of several distinct properties of rat lower motor neurons to isolate them from neighboring cells. First, an ideal stage in development after motor neurons are born (embryonic day 14 during rat gestation), but prior to extensive axonal extension or developmental apoptosis, is exploited. Lower motor neurons cannot be viably isolated using this method after birth. After dissociating embryonic spinal cord tissue, which contains lower motor neurons among many other cell types, the uniquely large motor neurons are enriched using density gradient centrifugation. Finally, the collected cell population is further purified based on selective immunopanning for motor neurons, which express the low-affinity nerve growth factor (NGF) receptor often referred to as p75. The near-pure lower motor neuron cultures are plated and seeded in defined conditions optimal for survival and can be maintained for several weeks. The expected yield is approximately 70,000 cells per embryonic spinal cord. |
| File Format | HTM / HTML |
| ISSN | 19403402 |
| Issue Number | 4 |
| Volume Number | 2013 |
| e-ISSN | 15596095 |
| Journal | Cold Spring Harbor Protocols |
| Language | English |
| Publisher | Cold Spring Harbor Laboratory Press |
| Publisher Date | 2013-04-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Clinical Laboratory Techniques Motor Neurons Physiology Animals Cell Culture Techniques Methods Cell Separation Cells, Cultured Embryo, Mammalian Rats Journal Article |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biochemistry, Genetics and Molecular Biology |
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