小鼠肾小管上皮细胞Cell Specification
Renal proximal tubular epithelial cells (RPTEpiC) play a crucial role in renal function. They
reabsorb nearly all the glucose and amino acids in the glomerular filtrate, while allowing
substances of no nutritional value to be excreted into the urine. They are also a major site of
injury in a variety of congenital, metabolic, and inflammatory diseases. RPTEpiC can produce
inflammatory mediators, such as cytokines or chemokines, and actively participate in acute
inflammatory processes by affecting and directing leukocyte chemotaxis via the production of
IL-8 [1, 2]. RPTEpiC express IL-2R alpha and MHC class II antigens during inflammation, after
renal transplantation, and during crescentic glomerulonephritis, suggesting that these cells have
the capacity to participate in the pathogenesis of immune renal injury [3]. To study the
relationship between proximal tubular cells and a variety of renal diseases, the RPTEpiC culture
is a useful in vitro model.
MRPTEpiC from ScienCell Research Laboratories are isolated from postnatal day 2 mouse
kidney. MRPTEpiC are cryopreserved at passage one and delivered frozen. Each vial contains
>5 x 105
cells in 1 ml volume. MRPTEpiC are characterized by immunofluorescence with
小鼠肾小管上皮细胞antibodies specific to cytokeratin-18, -19, and vimentin. MRPTEpiC are negative for
mycoplasma, bacteria, yeast, and fungi. MRPTEpiC are guaranteed to further culture under the
conditions provided by ScienCell Research Laboratories; however, MRPTEpiC are not
recommended for long term cultures due to limited expansion capacity and senescence after
subculturing.
Recommended Medium
It is recommended to use Epithelial Cell Medium-animal (EpiCM-a, Cat. #4131) for culturing
MRPTEpiC in vitro.
Product Use
MRPTEpiC are for research use only. They are not approved for human or animal use, or for
application in in vitro diagnostic procedures.
Storage
Upon receiving, directly and immediately transfer the cells from dry ice to liquid nitrogen, and
keep the cells in liquid nitrogen until they are needed for experiments.
Shipping
Dry ice.
References
小鼠肾小管上皮细胞[1] van Kooten C, van der Linde X, Woltman AM, van Es LA, Daha MR. (1999) “Synergistic effect of interleukin-1
and CD40L on the activation of human renal tubular epithelial cells.” Kidney Int. 56: 41-51.
[2] Schmouder RL, Strieter RM, Wiggins RC, Chensue SW, Kunkel SL. (1992) “In vitro and in vivo interleukin-8
production in human renal cortical epithelia.” Kidney Int. 41: 191-8.
[3] Wuthrich RP, Glimcher LH, Yui MA, Jevnikar AM, Dumas SE, Kelley VE. (1990) “MHC class II, antigen
presentation and tumor necrosis factor in renal tubular epithelial cells.” Kidney Int. 37: 783-92.
Instructions for culturing cells
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37oC water bath
and return the cells to culture as quickly as possible with minimal handling!
Initiating the culture:
1. Prepare a poly-L-lysine-coated culture vessel (2 μg/cm2
, T-75 flask is recommended).
Add 10 ml of sterile water to a T-75 flask and then add 15 μl of poly-L-lysine stock
solution (10 mg/ml, Cat. #0413). Leave the vessel in a 37oC incubator overnight (or for a
minimum of one hour).
2. Prepare complete medium. Decontaminate the external surfaces of medium bottle and
medium supplement tubes with 70% ethanol and transfer them to a sterile field.
Aseptically transfer supplement to the basal medium with a pipette. Rinse the supplement
tube with medium to recover the entire volume.
3. Rinse the poly-L-lysine-coated vessel twice with sterile water and then add 15 ml of
complete medium. Leave the vessel in the sterile field and proceed to thaw the
cryopreserved cells.
4. Place the frozen vial in a 37
oC water bath. Hold and rotate the vial gently until the
contents completely thaw. Promptly remove the vial from the water bath, wipe it down
with 70% ethanol, and transfer it to the sterile field.
小鼠肾小管上皮细胞5. Carefully remove the cap without touching the interior threads. Gently resuspend and
dispense the contents of the vial into the equilibrated, poly-L-lysine-coated culture vessel.
A seeding density of 5,000 cells/cm2
is recommended.
Note: Dilution and centrifugation of cells after thawing are not recommended since these
actions are more harmful to the cells than the effect of residual DMSO in the culture. It is
also important that cells are plated in poly lysine coated culture vessels to promote cell
attachment.
6. Replace the cap or lid of the culture vessel and gently rock the vessel to distribute the
cells evenly. Loosen cap if necessary to allow gas exchange.
7. Return the culture vessel to the incubator.
8. For best results, do not disturb the culture for at least 16 hours after the culture has been
initiated. Refresh culture medium the next day to remove residual DMSO and unattached
cells, then every other day thereafter.
Renal proximal epithelial cells are not recommended for long term culture due to
limited expansion capacity and senescence after subculturing
Caution: Handling animal derived products is potentially biohazardous. Always wear gloves
and safety glasses when working with these materials. Never mouth pipette. We recommend
小鼠肾小管上皮细胞following the universal procedures for handling products of human origin as the minimum
precaution against contamination [1].
[1] Grizzle WE, Polt S. (1988) “Guidelines to avoid personal contamination by infective agents in research
laboratories that use human tissues.” J Tissue Cult Methods. 11: 191‐9.