WORKFLOW KIT · STEM CELL THERAPY

HSC Enhancement Kit

Deliver functional enhancers into CD34+ hematopoietic stem cells, preserving engraftment potential and stemness.

Every kit includes: RNA · Optimized protocols · MicroBooster™ cartridges · Suggested readouts · Reagents

Upgrade your workflow

Boost a homing or survival mRNA into CD34+ HSCs, then transplant them the same day. Mechanoporation leaves the transcriptome close enough to untreated that the cells engraft without a recovery culture first, so the graft goes in while the mRNA is still being expressed.

The problem

Getting cargo into a CD34+ cell is not the hard part. Getting one back that still behaves like a stem cell is. Electroporation delivers, but the cells come out stressed, forming fewer colonies and engrafting worse. Protocols compensate with a recovery culture, which costs a day of transient expression before the graft goes in.

With this kit

Gateway™ mechanoporation transiently opens each CD34+ cell by mechanical deformation, so mRNA, CRISPR RNP, or protein diffuses straight into the cytosol before the cell reseals. Boosted cells track untreated controls on microarray, in the colony-forming assay, and in spleen and bone marrow chimerism, so they can go from boosting straight into the animal with no rest period.

Five steps, one boost

Gateway™ mechanoporation replaces the electroporation-plus-recovery step with a single, gentle delivery that the cells can be transplanted straight out of.

Portal has the Solution

Isolate

CD34+ HSCs

Mix

Cells + cargo (RNA / protein)

Boost

Gateway™ mechanoporation

Culture

HSC media post-boost

Confirm

Engraftment & viability

Unbiased, validated intracellular hits

>90%

Of human CD34+ HSCs express the boosted mRNA

3×

Spleen chimerism vs. electroporation, with no rest period before transplant

~80% edited

CD34+ HSCs after a single B2M CRISPR RNP boost, at >95% viability

What's in the box

The cargo and protocols below make this kit specific. The core consumables, settings, and support ship with every Portal kit.

Available Cargoes

CXCR4
BCL2

Don't see your cargo? Ask us about custom configurations.

Reagents & Consumables

Delivery tracer
MicroBooster™

Protocol & Settings

Protocol for HSC delivery
Recommended cell concentrations
Suggested assay readouts

Support

Portal RNA sourcing
Optional: hands-on implementation visit

Available Cargoes

CXCR4
BCL2

Don't see your cargo? Ask us about custom configurations.

Reagents & Consumables

Delivery tracer
MicroBooster™

Protocol & Settings

Protocol for HSC delivery
Recommended cell concentrations
Suggested assay readouts

Support

Portal RNA sourcing
Optional: hands-on implementation visit

Proof Points

Minimal Transcriptional Change in Boosted CD34+ Cells

Microarray on CD34+ HSCs at 6 and 24 hours after treatment, scoring genes misregulated more than 2-fold at p<0.05. Electroporated cells show broad up- and down-regulation at 6 hours that is still resolving at 24. Mechanoporated cells sit close to untreated controls at both timepoints, which is what lets the rest of the workflow skip a recovery step.

Engraftment Held at Control Levels With No Rest Period

CD34+ HSCs were injected into NBSGW mice at 100,000 cells per animal immediately after treatment, with no recovery culture in between, and read out at week 8. Mechanoporated cells gave about 13% human cells in the spleen, matching untreated control, against about 4.5% after electroporation. Bone marrow chimerism and CD14+ and CD19+ lineage output tracked control the same way, so both myeloid and lymphoid potential came through the boost intact.

Mutant CXCR4 Resists Plerixafor-Mediated Inhibition

CXCR4 directs an HSC to CXCL12 in the marrow niche, and Plerixafor blocks it. HSCs boosted with CXCR4 A175F mRNA kept CXCL12-induced Ca²⁺ flux nearly intact under Plerixafor, about 9% reduction against about 70% for Plerixafor alone and about 36% for wild-type CXCR4 mRNA. The boosted cells go on signaling under a drug that blunts signaling in the cells around them.

Mutant BCL2 Promotes HSC Survival Under Venetoclax

Venetoclax kills by inhibiting BCL2, and the G101V substitution stops the drug binding. CD34+ HSCs boosted with BCL2 G101V mRNA held about 87% of cells at 48 hours under Venetoclax, against about 19% for control and about 56% for wild-type BCL2. In NBSGW mice the same cargo raised transferred-cell frequency to about 62% of human cells under Venetoclax, against about 44% for a GFP control (p<0.0001), while the two were indistinguishable with no drug.

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mRNA, siRNA, saRNA

Proteins & Peptides

CRISPR RNPs

Small Molecules

Polymers

Nanoparticles

Antibodies

Virus

Validated cell types

PBMCs

T cells

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Monocytes

RBCs

HSCs

What is the HSC Enhancement Kit?The HSC Enhancement Kit boosts functional enhancer mRNA into primary human CD34+ hematopoietic stem cells in a single step, without a viral vector and without electroporation. Gateway™ mechanoporation transiently opens each cell by mechanical deformation, so mRNA reaches the cytosol directly and the cell reseals. The kit supplies CXCR4 and BCL2 mRNA, an HSC delivery protocol, recommended cell concentrations, and suggested readouts, and it is built for engraftment, survival, and stemness-preservation work in HSC gene therapy and transplant research.

What cargo can the HSC Enhancement Kit deliver to CD34+ cells?The HSC Enhancement Kit supplies CXCR4 and BCL2 mRNA, and the same Gateway™ delivery also carries CRISPR RNP, protein, and a dextran tracer into CD34+ HSCs. Portal's HSC work has boosted GFP, CXCR4, and BCL2 mRNA, an anti-B2M or anti-CCR5 CRISPR RNP, and both RNPs together in one delivery. Portal Bio sources the RNA as part of the kit and builds custom cargo configurations on request.

Does mechanoporation affect HSC engraftment?No. In NBSGW mice injected with 100,000 CD34+ cells per animal, mechanoporated HSCs gave about 13% human cells in the spleen at week 8, matching untreated control, while electroporated cells gave about 4.5%, roughly a 3-fold difference. Peripheral blood chimerism at weeks 4 to 10 and bone marrow chimerism at week 8 tracked control the same way, and so did lineage output, with the proportion of CD14+ myeloid and CD19+ lymphoid cells indistinguishable from untreated. Electroporation reduced both, most severely CD19+ lymphoid output.

Do HSCs need a rest period after mechanoporation before transplant?No. Mechanoporated CD34+ cells were transferred into NBSGW mice immediately after treatment with no recovery culture, and engrafted at untreated-control levels. Skipping the rest period is helpful for transient cargo because mRNA is expressed for hours to days, so a recovery culture spends part of that expression window before the cells reach the niche. Cells can also be plated into a colony-forming assay within 2 hours of the boost and still form colonies at control rates across CFU-GM, BFU-E, and CFU-GEMM.

Why does electroporation reduce HSC engraftment?Electroporation broadly misregulates the CD34+ transcriptome, and the functional readouts decline with it. Microarray at 6 hours after treatment shows hundreds of genes changed more than 2-fold at p<0.05, still resolving at 24 hours. Colony formation drops when cells are plated within 2 hours. In NBSGW mice, electroporated HSCs reached about 4.5% spleen chimerism against about 13% for untreated control at week 8, with reduced bone marrow chimerism and impaired CD19+ lymphoid output. Mechanoporated cells sit close to untreated controls on all three readouts.

What is CXCR4 A175F and why deliver it to HSCs?CXCR4 A175F is a Plerixafor-resistant variant of CXCR4, the chemokine receptor that directs a hematopoietic stem cell toward CXCL12 in the marrow niche, and it is supplied in the HSC Enhancement Kit as mRNA. Plerixafor is a CXCR4 antagonist used to mobilize HSCs, and the A175F substitution stops it inhibiting the receptor. HSCs boosted with CXCR4 A175F mRNA kept CXCL12-induced Ca²⁺ flux nearly intact under Plerixafor, about 9% reduction against about 70% for Plerixafor alone and about 36% for wild-type CXCR4 mRNA. Boosted CXCR4 also raised surface expression 4- to 6-fold in engrafted cells in bone marrow.

What is BCL2 G101V and why deliver it to HSCs?BCL2 G101V is a Venetoclax-resistant variant of the anti-apoptotic protein BCL2, supplied in the HSC Enhancement Kit as mRNA. Venetoclax kills cells by inhibiting BCL2, and the G101V substitution stops the drug binding. CD34+ HSCs boosted with BCL2 G101V mRNA held about 87% of cells at 48 hours under Venetoclax, against about 19% for control and about 56% for wild-type BCL2. In NBSGW mice the same cargo raised transferred-cell frequency to about 62% of human cells under Venetoclax against about 44% for a GFP control (p<0.0001), while the two groups were indistinguishable with no drug.

Ready to run the HSC Enhancement Kit?

Request the kit and a Portal scientist will spec it for your cells, cargo, and readout.

HSC Enhancement Kit

Upgrade your workflow

Five steps, one boost

What's in the box

Proof Points

Minimal Transcriptional Change in Boosted CD34+ Cells

Engraftment Held at Control Levels With No Rest Period

Mutant CXCR4 Resists Plerixafor-Mediated Inhibition

Mutant BCL2 Promotes HSC Survival Under Venetoclax

Check out our other kits

iPSC Generation

APC Enhancement

T Cell Functional Enhancement Kit

Neuronal Differentiation

More Cell Therapy kits

Portal’s platform supports delivery to a diverse range of cell types and cargoes

Frequently asked questions

Ready to run the HSC Enhancement Kit?