What makes the filtration process different from standard 0.22 micron filtered media?

FluxMPS runs the medium through four sequential filtration passes - 0.1 micron prefiltration I, 0.04 micron final filtration I, 0.1 micron prefiltration II, and 0.04 micron final filtration II - reaching a 0.04 micron final cut-off, five times finer than the 0.22 micron membranes used in conventional sterile filtration.

Does this medium require a CO2 incubator?

Yes, 5% CO2 is recommended. Sodium bicarbonate (2.0 g/L) provides the primary buffering system at this CO2 level; the 25 mM HEPES addition provides supplemental buffering capacity during bench handling outside the incubator.

FluxMPS RPMI 1640 Medium with 25mM HEPES w/o L-Glutamine, Phenol Red: 1X Liquid

verified ISO 13485 Certified Manufacturing

FluxMPS™ RPMI 1640 + 25mM HEPES

Sodium Bicarbonate HEPES (25 mM) Calcium Magnesium Glucose (2.0 g/L) Sodium Pyruvate L-Glutamine Phenol Red

FluxMPS™ DCP-RPMIH-QR1X is a Microfluidics Suitable, quadruple-stage ultra-filtered (0.1 µm ×2 + 0.04 µm ×2) RPMI 1640 + 25 mM HEPES formulation engineered for hematopoietic and lymphocyte-based cell models on organ-on-a-chip (OoC) and microphysiological system (MPS) platforms. The train reaches a 0.04 µm final cut-off — five times finer than the 0.22 µm membranes used for conventional sterile filtration. HEPES (25 mM, pKa 7.3 at 37°C) supplements the sodium bicarbonate buffering system for pH stability during bench handling and flow cytometry preparation. Formulation: [+] Sodium Bicarbonate, [+] HEPES (25 mM), [+] Calcium, [+] Magnesium, [+] Glucose (2.0 g/L), [+] Sodium Pyruvate | [-] L-Glutamine, [-] Phenol Red.

Glucose source: 2000 mg/L (2.0 g/L) D-glucose for standard glycolytic metabolism in RPMI-adapted cell lines
25 mM HEPES (pKa 7.3 at 37°C) supplements bicarbonate buffering for pH stability during bench handling and flow cytometry prep
Glutathione (reduced, 1.0 mg/L) included — antioxidant support for redox-sensitive lymphocyte and hematopoietic cell types
Quadruple-stage filtration train: 0.1 µm ×2 + 0.04 µm ×2, reaching a 0.04 µm final cut-off
Endotoxin release specification: < 0.05 EU/mL (LAL, USP <85>) — relevant for TLR4-sensitive immune cell assays
Phenol red–free and L-glutamine–free formulation — supports reduced-background imaging and researcher-controlled glutamine supplementation
Manufactured under an ISO 13485:2016 quality management system; final QC at Diagnocine, Totowa, NJ
pH 7.4; osmolality 280–320 mOsm/kg H₂O — formulated for RPMI-based lymphocyte and hematopoietic culture

CAT. NO.

DCP-RPMIH-QR1X | Cell Culture Media UNSPSC: 41116155 | Commodity: Molecular biology and cell culture growth media | (UNv260801)

RPMI 1640 + 25mM HEPES — 1X Liquid

Media familyRPMI 1640 + 25mM HEPES
Formulation[+] Sodium Bicarbonate, [+] HEPES 25mM, [+] Calcium, [+] Magnesium, [+] Glucose 2.0g/L, [+] Sodium Pyruvate | [-] L-Glutamine, [-] Phenol Red
AppearancePale yellow to colorless, clear solution
pH (USP <791>)7.4
Osmolality (USP <785>)280–320 mOsm/kg H₂O
Endotoxin (USP <85>)< 0.05 EU/mL
Filtration0.1 µm ×2 + 0.04 µm ×2 (Quadruple-stage)
Storage2–8°C, protect from light
Shelf Life12 months from date of manufacture, unopened
ShippingCold pack, temperature-monitored

ISO 13485:2016 USP <85> <785> <788> RUO

Why FluxMPS™

Engineered where standard media fails

Conventional 0.22 µm–filtered RPMI passes mycoplasma-scale organisms, subvisible particulates, and endotoxin fragments that can activate TLR4 signaling and confound immune cell assays. FluxMPS™ is built to reduce these risks at the filtration stage.

filter_alt

Microchannel-safe purity

0.04 µm final filtration with USP <788> Method 1 particulate compliance supports suspension immune cell OoC platforms and flow cytometry workflows sensitive to particulate interference.

target

Researcher-controlled metabolic inputs

L-glutamine is excluded, allowing precise, researcher-controlled nitrogen and glutamine-analog supplementation for T cell activation and metabolic flux studies. Sodium pyruvate (110 mg/L) and D-glucose (2.0 g/L) are provided as base carbon sources.

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Ultrapure-grade water

Formulated with Type 1 water (18.2 MΩ·cm) under trace-metal and organic-carbon control, reducing lot-to-lot variability contributed by feed water in sensitive lymphocyte culture systems.

visibility

Low background for imaging

Phenol red is excluded, reducing spectral interference for flow cytometry and confocal fluorescence microscopy. Riboflavin, present in all RPMI-based formulations, contributes intrinsic background fluorescence common to this medium class — plan filter sets accordingly.

science

Rich, stable nutrient profile

RPMI's 19-amino-acid and 10-vitamin profile, plus reduced glutathione for redox support, is optimized for lymphocyte, monocyte, and hematopoietic cell maintenance, released per-lot against the full specification matrix.

tune

Customization on demand

pH, nutrient concentrations, HEPES level, and component modifications available on request. Contact support@diagnocine.com.

Purity Architecture

Quadruple-stage filtration system

Four serial filtration passes — two dedicated 0.1 µm/0.04 µm prefilter-and-final-filter pairs — reach a final 0.04 µm polish under aseptic fill conditions.

1

0.1 µm Prefiltration I

Removes large particulate, cell debris, and protein aggregates; protects the first 0.04 µm cartridge.
2

0.04 µm Final filtration I — Mycoplasma Barrier

Retains organisms in the 0.2–0.3 µm mycoplasma size range — a pore size below what standard 0.22 µm filtration retains.
3

0.1 µm Prefiltration II

Second dedicated prefilter, protecting the second 0.04 µm final-filter cartridge.
4

0.04 µm Final filtration II — Polish

Ultimate polishing filter ahead of aseptic fill & finish in ISO Class 5 (Class 100) workstations.

Performance vs. conventional media

A quadruple-stage train (0.1 µm ×2 + 0.04 µm ×2) reaches a 0.04 µm final cut-off, five times finer than the 0.22 µm membranes used for conventional sterile filtration.

4

Sequential filtration passes: 0.1 µm ×2 + 0.04 µm ×2

0.04 µm

Final pore size — 5.5× finer than standard 0.22 µm filtration

Sterility: No growth after 14-day incubation (USP <71>). Mycoplasma: controlled by 0.1 µm mycoplasma-retentive filtration (not tested per lot).

Grade: This product is Microfluidics Suitable, filtered to a 0.04 µm final cut-off. It is not an MPS Grade product — that designation is reserved for the 0.01 µm ultra nano-filtered line, which adds 0.02 µm and 0.01 µm stages after the 0.04 µm polish. For applications requiring the 0.01 µm cut-off, contact support@diagnocine.com.

FluxMPS DCP-RPMIH-QR1X RPMI 1640 25mM HEPES quadruple-stage filtration system 0.1 micron x2 plus 0.04 micron x2 for organ-on-a-chip and microfluidic cell culture, Diagnocine — **Figure 1.** FluxMPS™ Quadruple-stage filtration system (0.1 µm ×2 + 0.04 µm ×2).
© Diagnocine® — DCP-RPMIH-QR1X

Applications

Immune cell OoC and hematopoietic applications

FluxMPS™ DCP-RPMIH-QR1X — RPMI 1640 + 25mM HEPES — delivers 0.04 µm filtered purity for hematopoietic cells and related OoC applications.

Automated Bioreactors & Robotics

Next-Generation System Uptime

An optional 0.01 µm (10 nm) MPS Grade variant of this formulation — a separate six-stage ultra nano-filtered line beyond the Microfluidics Suitable 0.04 µm tier described above — is available on request for automated bioreactor and robotic perfusion systems.

Total Particulate Exclusion: 0.01 µm filtration targets nanoparticulate aggregates beyond the 0.04 µm tier
Valve & Sensor Protection: Reduces micro-fouling risk in automated perfusion systems
Extended Perfusion Stability: Consistent nutrient delivery over long-duration culture

Inquiry Required: Contact support@diagnocine.com for the 0.01 µm MPS Grade variant.

Immunology

T Cell & Lymphocyte Culture

RPMI 1640 is the standard base for primary T cells, B cells, NK cells, and monocytes. FluxMPS™ 0.04 µm filtration reduces particulate load that can confound immune assays.

T cellsB cellsNK cellsPBMC

Cancer Biology

Leukemia & Lymphoma Lines

RPMI supports NCI-60 hematologic cancer lines, Jurkat, Raji, K562, and HL-60, where DMEM would alter proliferation and signaling behavior.

JurkatRajiK562HL-60

Microfluidics

Immune Cell OoC

0.04 µm filtered RPMI supports tumor-immune interaction chips, vascular-immune OoC, and lymph node-on-chip models sensitive to particulate load.

Tumor-immune chipLymph node OoCMPS

Immunotherapy

CAR-T & TIL Expansion

The <0.05 EU/mL endotoxin release specification supports CAR-T manufacturing and TIL expansion protocols where endotoxin-driven T cell activation artefacts are a concern.

CAR-TTILTCR-T

Metabolomics

Immune Cell Metabolic Flux

Defined RPMI base for glycolysis stress assays and ¹³C isotope tracing of lymphocyte activation states. Not compatible with Agilent Seahorse XF assays, which require bicarbonate-free, phenol red–free medium.

¹³C tracingGlycolysis stress testLC-MS metabolomics

Live-Cell Imaging

Flow Cytometry & Confocal

Ultra-low particulate and phenol red–free formulation reduce spectral background for PE-channel flow cytometry and immune cell confocal imaging.

Flow cytometryConfocalELISA

Technical Specifications

Analytical release specifications

Every lot released against the full specification matrix. CoA: support@diagnocine.com.

Physical & Chemical Parameters

Parameter	Specification
Formulation	[+] Sodium Bicarbonate, [+] HEPES 25mM, [+] Calcium, [+] Magnesium, [+] Glucose 2.0 g/L, [+] Sodium Pyruvate \| [-] L-Glutamine, [-] Phenol Red
Appearance	Pale yellow to colorless, clear solution
Glucose	2000 mg/L (2.0 g/L)
HEPES	25 mM (pKa 7.3 at 37°C)
pH USP <791>	7.4
Osmolality USP <785>	280–320 mOsm/kg H₂O
Total ingredients	40 components across 3 categories

Sterility, Purity & Safety

Parameter	Specification
Endotoxin USP <85> BET	< 0.05 EU/mL (batch release spec, see Manufacturing & Compliance)
Sterility USP <71>	No growth / 14 days
Mycoplasma	0.1 µm mycoplasma-retentive filtration (not tested per lot)
Particulate ≥10 µm USP <788> Method 1	NMT 25/mL
Particulate ≥25 µm USP <788> Method 1	NMT 3/mL
Water purity	Type 1, 18.2 MΩ·cm
Manufacturing std.	ISO 13485:2016
Fill environment	ISO Class 5 (Class 100)

Storage, Handling & Logistics

Parameter	Specification
Storage temperature	2–8°C, away from light
Freeze-thaw	Do not freeze
Shelf life	12 months from date of manufacture, unopened
Shipping condition	Cold pack, temperature-monitored
CO₂ requirement	5% CO₂ recommended; sodium bicarbonate (2.0 g/L) is the primary buffer, with 25 mM HEPES as supplemental buffering during bench handling

Raw Materials & Regulatory

Parameter	Specification
Raw material grade	Reagent / cell culture grade
Traceability	Full lot traceability per ISO 13485
Manufacturing QMS ISO	ISO 13485:2016 certified
UNSPSC	41116155 — Molecular biology and cell culture growth media (UNv260801)
Grade	Microfluidics Suitable (0.04 µm final cut-off)
Regulatory alignment	21 CFR Part 820 (QMSR) aligned
Production method	Micro-batch, per-lot QC release
Intended use	Research Use Only (RUO)

Formulation

Full composition (mg/L)

RPMI 1640 + 25mM HEPES: 40 ingredients verified per lot with CAS numbers for raw-material traceability. This formulation includes reduced glutathione and a balanced amino acid and vitamin profile optimized for lymphocyte and hematopoietic cell culture.

Component	CAS Number	mg/L
INORGANIC SALTS
Calcium nitrate tetrahydrate	13477-34-4	100.000
Magnesium sulfate anhydrous	7487-88-9	48.840
Potassium chloride	7447-40-7	400.000
Sodium bicarbonate	144-55-8	2000.000
Sodium chloride	7647-14-5	6000.00
Sodium phosphate dibasic anhydrous	7558-79-4	800.000

Component	CAS Number	mg/L
AMINO ACIDS
Glycine	56-40-6	10.000
L-Arginine hydrochloride	1119-34-2	241.000
L-Asparagine	70-47-3	50.000
L-Aspartic acid	56-84-8	20.000
L-Cystine dihydrochloride	30925-07-6	65.200
L-Glutamic acid	56-86-0	20.000
L-Histidine hydrochloride monohydrate	5934-29-2	20.960
L-Hydroxyproline	51-35-4	20.000
L-Isoleucine	73-32-5	50.000
L-Leucine	61-90-5	50.000
L-Lysine hydrochloride	657-27-2	40.000
L-Methionine	63-68-3	15.000
L-Phenylalanine	63-91-2	15.000
L-Proline	147-85-3	20.000
L-Serine	56-45-1	30.000
L-Threonine	72-19-5	20.000
L-Tryptophan	73-22-3	5.000
L-Tyrosine Disodium Salt	69847-45-6	28.830
L-Valine	72-18-4	20.000

Component	CAS Number	mg/L
VITAMINS
Choline chloride	67-48-1	3.000
D-Biotin	58-85-5	0.200
D-Ca-Pantothenate	137-08-6	0.250
Folic acid	59-30-3	1.000
Niacinamide	98-92-0	1.000
Pyridoxine hydrochloride	58-56-0	1.000
Riboflavin	83-88-5	0.200
Thiamine hydrochloride	67-03-8	1.000
Vitamin B12	68-19-9	0.005
p-Amino benzoic acid (PABA)	150-13-0	1.000
OTHERS
D-Glucose	50-99-7	2000.000
Glutathione reduced	70-18-8	1.000
HEPES	7365-45-9	5958.00
Sodium Pyruvate	113-24-6	110.000
i-Inositol	87-89-8	35.000

Custom formulation: Contact support@diagnocine.com for DCP-RPMIH-QR1X modifications.

Quality Assurance

Manufacturing & compliance

Every FluxMPS™ product is manufactured and released under a multi-layer quality system.

verified

ISO 13485:2016 Quality Management

Manufactured under an ISO 13485:2016-certified quality management system. Final QC at Diagnocine's R&D Center, Totowa, NJ, USA.

water_drop

Ultrapure Type 1 Water

18.2 MΩ·cm Type 1 water processed under trace-metal and organic-carbon control.

biotech

ISO Class 5 Fill & Finish

Aseptic fill in validated ISO Class 5 (Class 100) laminar-flow workstations.

assignment

Micro-Batch Precision

Small-batch, per-lot tested — no blending; Certificate of Analysis issued for every lot.

Endotoxin — USP <85> BET

LAL assay; release specification < 0.05 EU/mL per batch.

Particulate — USP <788> Method 1

≤25/mL (≥10 µm), ≤3/mL (≥25 µm), light obscuration.

Osmolality — USP <785>

Target: 280–320 mOsm/kg H₂O.

Documentation & CoA

Full CoA with raw-material traceability available on request.

Batch-level quality control. Endotoxin is controlled per manufacturing batch rather than per unit. Every batch is tested before release and must meet the release specification:

Endotoxin — LAL assay, USP <85> Bacterial Endotoxins Test; assay sensitivity 0.005 EU/mL; release specification < 0.05 EU/mL
pH, osmolality, conductivity, appearance and clarity
Sterility

A Certificate of Analysis is available on request at support@diagnocine.com.

Product Comparison

How DCP-RPMIH-QR1X compares

FluxMPS™ DCP-RPMIH-QR1X vs. conventional 0.22 µm–filtered RPMI formulations.

Parameter	DCP-RPMIH-QR1X (FluxMPS™)	Conventional RPMI 1640 + HEPES (0.22 µm filtered)	Standard DMEM/RPMI (0.22 µm filtered)
Grade	Microfluidics Suitable	Not specified	Not specified
RPMI 1640 + 25mM HEPES — no L-Glutamine, no Phenol Red; pH-stable imaging-clean immune base	check_circle Yes	cancel No	cancel No
Final filtration pore size	0.04 µm	0.22 µm	0.22 µm
Number of filtration stages	4 (Quadruple)	1	1
Mycoplasma-retentive filtration	check_circle Yes (0.1 µm stage)	cancel No	cancel No
Endotoxin (release specification)	FluxMPS™ — < 0.05 EU/mL	Corning classical liquid media — < 0.25 EU/mL Sigma-Aldrich DMEM complete medium — ≤ 2 EU/mL Gibco classical DMEM — Not specified (recorded per lot)
USP <788> particulate tested	check_circle Yes	cancel No	cancel No
Water quality	Type 1, 18.2 MΩ·cm	Purified water	Purified water
Manufacturing QMS	ISO 13485:2016	ISO 9001 or none	ISO 9001 or none
Microfluidic channel compatibility	check_circle Yes (Microfluidics Suitable)	cancel Risk of clogging	cancel Risk of clogging
Custom formulation available	check_circle Yes	cancel No	cancel No

Comparison figures from published supplier specifications, accessed 2026-09-02. Suppliers that publish no numeric endotoxin specification are shown as "Not specified".

FAQ

Frequently asked questions

Common questions about FluxMPS™ DCP-RPMIH-QR1X — RPMI 1640 + 25mM HEPES.

Is DCP-RPMIH-QR1X suitable for organ-on-a-chip (OoC) applications?expand_more

Yes. DCP-RPMIH-QR1X is processed through a quadruple-stage filtration system (0.1 µm ×2 + 0.04 µm ×2) reaching a 0.04 µm final cut-off, well suited to immune cell OoC, tumor-immune interaction chips, and lymphocyte perfusion models where standard 0.22 µm–filtered RPMI can introduce particulate and mycoplasma-scale contaminants.

What makes the filtration process different from standard 0.22 µm filtered media?expand_more

FluxMPS™ runs the medium through four sequential filtration passes — 0.1 µm prefiltration I, 0.04 µm final filtration I, 0.1 µm prefiltration II, and 0.04 µm final filtration II — reaching a 0.04 µm final cut-off, five times finer than the 0.22 µm membranes used in conventional sterile filtration.

Why are L-glutamine and phenol red both excluded, and how do I supplement?expand_more

Phenol red is excluded to reduce spectral background for flow cytometry and fluorescence imaging. L-glutamine is excluded because it degrades in liquid storage; add fresh L-glutamine or a stable dipeptide substitute at the concentration required by your cell line at the time of use. Sodium pyruvate (110 mg/L) and D-glucose (2.0 g/L) are already included as base carbon sources.

Does this medium require a CO₂ incubator?expand_more

Yes, 5% CO₂ is recommended. Sodium bicarbonate (2.0 g/L) provides the primary buffering system at this CO₂ level; the 25 mM HEPES addition provides supplemental buffering capacity during bench handling outside the incubator.

Can DCP-RPMIH-QR1X be supplemented with serum, growth factors, or other components?expand_more

Yes. FBS (5–10%), serum-free supplements, growth factors, and antibiotics can be added. When filtering serum-containing additions, use a 0.2 µm low-protein-binding PES or PVDF filter — never a 0.04 µm membrane, which retains immunoglobulins and lipoproteins present in serum. Contact support@diagnocine.com for custom co-formulation.

What endotoxin level should I expect, and how is it verified?expand_more

The release specification is less than 0.05 EU/mL, verified by LAL assay per USP <85> (Bacterial Endotoxins Test; assay sensitivity 0.005 EU/mL). Endotoxin is controlled per manufacturing batch: every batch is tested before release and must meet this specification before shipment.

Is a Certificate of Analysis (CoA) available, and what does it contain?expand_more

Yes. Each lot's CoA covers appearance, pH (USP <791>), osmolality (USP <785>), sterility (USP <71>), endotoxin (USP <85>), mycoplasma-retentive filtration status, and particulate count (USP <788> Method 1). Request at support@diagnocine.com.

Scientific References

Supporting literature

Key publications supporting RPMI 1640 + 25mM HEPES in immune cell culture and OoC applications.

Moore GE, et al. Culture of normal human leukocytes. JAMA. 1967;199:519–524. doi:10.1001/jama.1967.03120080053007
Huh D, et al. Reconstituting organ-level lung functions on a chip. Science. 2010;328:1662–1668. doi:10.1126/science.1188302
Bhatia SN, Ingber DE. Microfluidic organs-on-chips. Nat Biotechnol. 2014;32:760–772. doi:10.1038/nbt.2989
Novak R, et al. Robotic fluidic coupling and interrogation of multiple vascularized organ chips. Nat Biomed Eng. 2020;4:407–420. doi:10.1038/s41551-019-0497-x
Jang KJ, et al. Human kidney proximal tubule-on-a-chip. Integr Biol. 2013;5:1119–1129. doi:10.1039/c3ib40049b
Schimek K, et al. Integrating biological vasculature into a multi-organ-chip microsystem. Lab Chip. 2013;13:3588–3598. doi:10.1039/c3lc50217a
Boussommier-Calleja A, et al. Microfluidics: A new tool for modeling cancer-immune interactions. Trends Cancer. 2016;2:6–19. doi:10.1016/j.trecan.2015.12.003
Sung JH, et al. Microfabricated mammalian organ systems. Lab Chip. 2013;13:1201–1212. doi:10.1039/c3lc41017j

FluxMPS™ RPMI 1640 Medium with 25mM HEPES w/o L-Glutamine, Phenol Red: 1X Liquid