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大鼠嗜碱性粒细胞性白血病细胞 RBL-1
科研骨髓间充质干细胞（BMSC）实验用
通过广州市番禺区工商行政管理局认证 
Corning 手动单道/多
Corning 通用离心机
康宁陶瓷搅拌加热板 C
Corning 康宁精密水浴
FIX&PERM® Sampl
FIX&PERM® Cell 
FIX&PERM® Solut
FIX&PERM® Solut| 货号： | HSC001 |
| 生长状态： | 存活率95%以上 |
| 年限： | 6个月 |
| 运输方式： | Liquid Nitrogen |
| 器官来源： | Liver |
| 是否是肿瘤细胞： | NO |
| 细胞形态： | 多边，贴壁 |
| 免疫类型： | NC |
| 物种来源： | Mice/rat/human |
| 相关疾病： | NC |
| 组织来源： | Liver |
| 品系： | NC |
| ATCC Number： | NC |
| 细胞类型： | BIOCYTO |
| 供应商： | BIOCYTO |
| 规格： | 10^6 |
| Product | Catalog no. | Amount | Storage |
| Hepatic Stellate | HSC001 | 1X10^6/vial | in liquid nitrogen |
Product Use
For Research Use Only.Not for use in diagnostic procedures
Culture Conditions
Culture Type:Adherent
Temperature Range:36℃ to 38℃
Incubator Atmosphere:Humidified atmosphere of 5% CO2
Size:The typical hepatocyte is cubical with sides of 20-30 µm
Passaging Adherent Cells
All solutions and equipment that come in contact with the cells must be sterile.
Always use proper sterile technique and work in a laminar flow hood.
1. Remove and discard the spent cell culture media from the culture vessel.
2. Wash cells using a balanced salt solution without calcium and magnesium (approximately 2 mL per 10 cm2 culture surface area). Gently add washsolution to the side of the vessel opposite the attached cell layer to avoiddisturbing the cell layer,and rock the vessel back and forth several times.
Note: The wash step removes any traces of serum, calcium, and magnesium that would inhibit the action of the dissociation reagent.
3. Remove and discard the wash solution from the culture vessel
4. Add the pre-warmed dissociation reagent such as trypsin or TrypLE™to the side of the flask; use enough reagent to cover the cell layer (approximately0.5 mL per 10 cm2). Gently rock the container to get complete coverage of the cell layer.
5. Incubate the culture vessel at room temperature for approximately 2minutes.
Note: that the actual incubation time varies with the cell line used.
6. Observe the cells under the microscope for detachment. If cells areless than 90% detached, increase the incubation time a few more minutes, checking for dissociation every 30 seconds. You may also tap the vessel to expedite cell Tetachment.
7. When ≥ 90% of the cells have detached, tilt the vessel for a minimallength of time to allow the cells to drain. Add the equivalent of 2 volumes (twice thevolume used for the dissociation reagent) of pre-warmed complete growth medium.Disperse the medium by pipetting over the cell layer surface several times.
8. Transfer the cells to a 15-mL conical tube and centrifuge then at200 × g for 5 to 10 minutes. Note that the centrifuge speed and time vary based on the cell type.
9. Resuspend the cell pellet in a minimal volume of pre-warmed complete growth medium and remove a sample for counting.
10. Determine the total number of cells and percent viability using a hemacytometer, cell counter and Trypan Blue exclusion, or the Countess® Automated CellCounter. If necessary, add growth media to the cells to achieve the desired cellconcentration and recount the cells.
Hepatic stellate cells (here HSC), also known as perisinusoidal cells or Ito cells (earlier lipocytes or fat-storing cells), are pericytes found in the perisinusoidal space (a small area between the sinusoids and hepatocytes) of the liver also known as the space of Disse. The stellate cell is the major cell type involved in liver fibrosis, which is the formation of scar tissue in response to liver damage.
Function
In normal liver, stellate cells are described as being in a quiescent state. Quiescent stellate cells represent 5-8% of the total number of liver cells. Each cell has several long protrusions that extend from the cell body and wrap around the sinusoids. The lipid droplets in the cell body store vitamin A as retinol ester. The function and role of quiescent hepatic stellate cells is unclear. Recent evidence suggests a role as a liver-resident antigen-presenting cell, presenting lipid antigens to and stimulating proliferation of NKT cells.
When the liver is damaged, stellate cells can change into an activated state. The activated stellate cell is characterized by proliferation, contractility, and chemotaxis. The amount of stored vitamin A decreases progressively in liver injury. The activated stellate cell is also responsible for secreting collagen scar tissue, which can lead to cirrhosis. More recent studies have also shown that in vivo activation of hepatic stellate cells by agents causing liver fibrosis can eventually lead to senescence of these cells, marked by increased SA-beta-galactosidase staining, as well as p53 accumulation and activation of Rb–hallmarks of cellular senescence. The senescent hepatic stellate cells have been demonstrated to limit liver fibrosis by activating the immune system by activating interactions with the NK cells.
