R10 Medium (RPMI + 10% FCS + Pen/Strep)
Dillon Corvino
Complete RPMI 1640 medium supplemented with 10% fetal calf serum and Penicillin/Streptomycin for culture and washing of human PBMCs and lymphocytes.
Protocol ID: BUF-R10-MED Version: v1.1 Author: Dillon Corvino Date: September 18, 2026
Purpose
R10 is complete RPMI 1640 medium supplemented with 10% heat-inactivated FCS and Penicillin/Streptomycin. It is used for PBMC washing after Ficoll separation, for quenching CellTrace dyes, and as a general-purpose lymphocyte culture medium.
Linked protocols
HUM-BLD-PBMC-ISOHUM-KHYG1-CULTUREGEN-CTV-STAIN
Working buffer composition
| Component | Final concentration | Notes |
|---|---|---|
| RPMI 1640 | — | Base medium |
| Fetal calf serum | 10% (v/v) | Heat-inactivated |
| Penicillin/Streptomycin | 1% (v/v) | 100x stock → 1x final |
Preparation
A. Preparation of stock (if applicable)
Not applicable — prepared directly as the working buffer.
B. Preparation of working buffer
Stock solutions required
| Reagent | Stock concentration | Notes |
|---|---|---|
| Fetal calf serum (FCS) | — | Heat-inactivate if required |
| Penicillin/Streptomycin | 100x | Add to 1x final |
| RPMI 1640 | — | Sterile-filtered |
Preparation of 500 mL R10
- Aliquot 450 mL sterile RPMI 1640 into a sterile bottle.
- Add 50 mL FCS (10% final).
- Add 5 mL Penicillin/Streptomycin (100x stock) → 1% final.
- Gently invert to mix; do not vortex.
- Label with:
- R10 complete medium
- Date of preparation
- Expiry (4 weeks after preparation)
Tip
Pre-warming R10 to 37 °C before use improves cell recovery following Ficoll separation or staining.
Storage and stability
- Store at 4 °C, protected from light.
- Use within 4 weeks after preparation.
- Do not freeze.
Reagent details
| Reagent | Supplier | Cat. # | Notes |
|---|---|---|---|
| RPMI 1640 | Various | — | Base medium |
| Fetal calf serum | Various | — | Heat-inactivated |
| Pen/Strep (100x) | Various | — | Final 1x concentration |
Safety (brief)
- Handle medium aseptically in a biosafety cabinet.
- Dispose of contaminated or expired medium as biohazardous waste.
- Follow institutional guidelines for handling FCS-containing media.
Version history
| Version | Date | Author | Changes |
|---|---|---|---|
| v1.0 | 2025-11-21 | Dillon Corvino | Initial R10 buffer document. |
| v1.1 | 2026-09-18 | Dillon Corvino | Migrated to shared summary:/helper architecture (buffer/protocol redesign). |