Rabbit Primary Cardiac Microvascular Endothelial Cells

Description

Establishing a primary cardiac microvascular endothelial cell culture requires clear confirmation of tissue origin, endothelial identity, passage, microbial quality, and post-arrival handling conditions. Rabbit Primary Cardiac Microvascular Endothelial Cells, Catalog No. N-6024, provide a documented source of primary rabbit cardiac endothelial cells isolated from the heart tissue of New Zealand White Rabbits.

The cells are grown in gelatin pre-coated tissue culture flasks with Complete Growth Medium. Cells at passage 3 are harvested and cryopreserved in vials at a documented concentration of 0.5 × 10⁶ cells per mL.

These rabbit cardiac microvascular endothelial cells are characterized by uptake of fluorescence-labeled acetylated low-density lipoprotein (DiI-Ac-LDL), a functional marker for endothelial cells. Product documentation reports negative results for bacteria, yeast, fungi, and mycoplasma.

Under the specified primary endothelial cell culture conditions, the cells may be expanded for 3-5 passages using a recommended split ratio of 1:2. Repeated freezing and thawing is not recommended.

For Research Use Only: This product is not authorized for human use, in vitro diagnostic procedures, or therapeutic procedures.

Specifications

The product datasheet documents the cryopreserved concentration as 0.5 × 10⁶ cells per mL. It does not specify the total cryovial volume.

Features

• Primary rabbit cardiac microvascular endothelial cells isolated from New Zealand White Rabbit heart tissue
• Harvested and cryopreserved at passage 3
• Documented cryopreserved concentration of 0.5 × 10⁶ cells/mL
• Characterized by DiI-Ac-LDL uptake as a functional endothelial cell marker
• Negative for bacteria, yeast, fungi, and mycoplasma
• Expandable for 3-5 passages under the specified culture conditions
• Recommended split ratio of 1:2
• Cultured on gelatin-coated cultureware at 37°C with 5% CO₂
• Cryopreserved cells are shipped on dry ice and require immediate liquid nitrogen storage

Ordering Formats

The supplied documentation does not specify a fixed cell count for the T25 or T75 live-cell formats. Confirm live-cell density, flask condition, and shipment scheduling before ordering when these details are required for experimental planning.

Applications

N-6024 supports research workflows requiring primary rabbit cardiac microvascular endothelial cell culture. The supplied protocol provides procedures for recovering cryopreserved cells, maintaining adherent endothelial cultures, expanding cells, preparing cultures for cell-based assays, and preserving cells for future research.

• Primary rabbit cardiac microvascular endothelial cell culture
• Primary endothelial cell recovery and expansion
• Cell-based assays using cultured primary endothelial cells
• Cell staining workflows
• FACS sample preparation
• Light-microscopy monitoring of cell confluence and morphology
• Primary cell subculture and cryopreservation

Researchers should independently determine whether N-6024 cells and the documented culture conditions are appropriate for their intended experimental design. The supplied documentation does not provide product-specific validation for disease modeling, angiogenesis, drug screening, or barrier-function assays.

Cell Culture Workflow

Culture Preparation

• Perform all cell culture procedures in a biosafety cabinet using aseptic technique.
• Sterile filter culture media, supplements, and reagents through a 0.2 µm filter.
• Prewarm only the volume of medium needed for immediate use to 37°C.
• Use 5 mL of medium for a T25 flask and 8-10 mL for a T75 flask when recovering cells, changing medium, or splitting cells.
• Repeated prewarming of complete culture medium is not recommended.

Gelatin Coating

• Coat sterile culture dishes or flasks with Gelatin-Based Coating Solution, Catalog No. CB6950, for 2 minutes.
• Aspirate the excess coating solution before seeding cells.

Recovery from a Cryovial

1. Thaw the vial rapidly in a 37°C water bath for less than 1 minute, or until only a few ice pieces remain.
2. Remove the vial immediately and disinfect its exterior with 70% ethanol.
3. Transfer the cells into a sterile 15 mL conical tube containing 5 mL of prewarmed cell culture medium.
4. Rinse the vial with an additional 0.5-1 mL of medium to collect the remaining cells.
5. Centrifuge at 200 × g for 5 minutes to remove residual DMSO.
6. Carefully aspirate the supernatant and resuspend the cell pellet in 5 mL of Complete Growth Medium.
7. Seed the resuspended cells into a T25 flask pre-coated with gelatin-based coating solution.
8. Culture for 12-16 hours in a humidified incubator at 37°C with 5% CO₂.
9. Change the medium on the second day and every 24 hours thereafter.
10. Check confluence and appropriate cell morphology daily under a microscope.

Primary Cell Expansion

1. Remove the culture medium and wash the adherent cells twice with room-temperature 1X PBS without calcium or magnesium.
2. Incubate cells with prewarmed 0.25% Trypsin-EDTA, Catalog No. CB6914, for 3-5 minutes.
3. Use 0.5-1 mL of Trypsin-EDTA for a T25 flask or 2 mL for a T75 flask.
4. Once the cells detach, immediately add 5-10 mL of complete culture medium containing fetal bovine serum to neutralize the trypsin.
5. Gently pipette the cell suspension up and down several times.
6. Seed cells into fresh gelatin-coated flasks or plates at the recommended split ratio of 1:2.
7. Return the cultures to a humidified 37°C, 5% CO₂ incubator.
8. Change the medium on the second day and every 24 hours thereafter.

Preparation for Experiments

• Cells should be checked daily for confluence and appropriate morphology.
• Once confluent, cultures may be allowed to overgrow for 24 hours before cell-based assays, staining, FACS analysis, or other designed experiments.
• Change the medium daily and wash the cells twice with 1X PBS before each medium change.
• Before an experiment, the protocol recommends using a 40 µm cell strainer to remove cell clumps and most dead or floating large cells.

Cryopreservation

• Detach cells with 0.25% Trypsin-EDTA and centrifuge at 200 × g for 5 minutes.
• Resuspend the cell pellet in 1 mL of Freezing Buffer, Catalog No. CB6916, per vial.
• The documented freezing buffer formulation is 10% DMSO, 50% FBS, and 40% culture medium.
• Place vials in a controlled-rate freezing container filled with 100% isopropanol at -80°C for 6-12 hours.
• Transfer the vials to liquid nitrogen for long-term storage.

The temporary -80°C step applies only to the documented laboratory cryopreservation procedure. Received cryopreserved cells must be transferred directly from the dry-ice shipping container to liquid nitrogen upon arrival.

Recommended Materials

These materials are referenced by the product datasheet or cell culture protocol. The documentation does not state that they are included with N-6024.

Quality Control

Storage and Handling

Immediate storage required: Transfer cryopreserved cells from the dry-ice shipping container to liquid nitrogen at -180°C immediately upon arrival. Do not use a -20°C or -80°C freezer for routine storage of the received primary cells.

• Cryopreserved cells are shipped overnight on dry ice.
• Live-cell shipment is available on request.
• Repeated freezing and thawing is not recommended.
• Handle all animal cells with caution and treat them as potential pathogens.
• No testing procedure can completely guarantee the absence of infectious agents.
• The cells are intended only for research use.

Warranty and Use Conditions

The manufacturer documentation states that its guarantee applies only when Cell Biologics cells are purchased with the specified Cell Biologics medium and coating solution, cultured and tested according to the applicable protocol, and evaluated within 35 days of product delivery.

Transfer or resale of Cell Biologics cells or products to other markets, organizations, or individuals is prohibited without the manufacturer's written consent. Applicable terms and conditions must be accepted before ordering.

Documents

N-6024 Product Datasheet

Primary Cell Culture Protocol

Frequently Asked Questions

What are Rabbit Primary Cardiac Microvascular Endothelial Cells?

They are primary cardiac microvascular endothelial cells isolated from the heart tissue of New Zealand White Rabbits. N-6024 cells are harvested and cryopreserved at passage 3.

What is the tissue source of N-6024 cells?

N-6024 rabbit cardiac endothelial cells are isolated from New Zealand White Rabbit heart tissue.

What is the concentration of the cryopreserved cells?

The product datasheet documents a cryopreserved cell concentration of 0.5 × 10⁶ cells per mL. The datasheet does not state the total cryovial volume, so this value should not be interpreted as the total number of cells per vial.

How are the rabbit cardiac microvascular endothelial cells characterized?

The cells are characterized by uptake of fluorescence-labeled acetylated low-density lipoprotein, or DiI-Ac-LDL. The datasheet identifies DiI-Ac-LDL uptake as a functional marker for endothelial cells.

What microbial quality testing is documented?

The product documentation reports negative results for bacteria, yeast, fungi, and mycoplasma.

How many passages can N-6024 cells be expanded?

Under the specified primary endothelial cell culture conditions, the cells may be expanded for 3-5 passages using a recommended split ratio of 1:2.

What medium is recommended for rabbit endothelial cell culture?

The N-6024 product datasheet recommends H1168 Complete Endothelial Cell Medium with Kit - 500 mL.

Do culture flasks require coating before seeding?

Yes. The supplied protocol specifies coating sterile culture dishes or flasks with CB6950 Gelatin-Based Coating Solution for 2 minutes and aspirating the excess solution before seeding cells.

What are the recommended incubation conditions?

The documented primary cell culture condition is a humidified incubator at 37°C with 5% CO₂.

How should cryopreserved cells be handled upon arrival?

Transfer the cells immediately from the dry-ice shipping container to liquid nitrogen at -180°C. Received primary cells should not be kept in a -20°C or -80°C freezer for routine storage.

What is the difference between the frozen vial, T25 flask, and T75 flask formats?

The frozen vial contains cryopreserved cells and is shipped on dry ice. T25 and T75 flasks are live-cell formats. The supplied documentation does not specify fixed cell counts for the live-cell flask formats, so cell density and shipment condition should be confirmed before ordering when required for experimental planning.

Can the cells be repeatedly frozen and thawed?

No. Repeated freezing and thawing of primary cells is not recommended.

Are N-6024 cells intended for clinical, diagnostic, or therapeutic use?

No. N-6024 cells are for research use only and are not authorized for human use, in vitro diagnostic procedures, or therapeutic procedures.