In 2018, at the Cambridge International Stem Cell Symposium, we presented the results of our self-made serum-free medium (HELENE Medium). First, HELENE Medium allows more cells to be cultured in the same period of time than other commercially available media, and second, the mesenchymal stem cells cultured in HELENE Medium are the smallest in size. There is scientific evidence that smaller stem cells are less likely to differentiate into fibroblasts.
Combining the above two points, it was confirmed that the adipose tissue collected from behind the ear contains MSCs. The amount of fat collected is small compared to thighs and abdomens, but with our unique laboratory technology and HELENE Medium, it is possible to culture up to 2.25 billion MSCs in one month.
In order to guarantee the quality of the stem cells we provide to our patients, we have outsourced testing of stem cell samples manufactured in the same process as this quality certificate. The test complies with GMP (Good Manufacturing Practice) and regenerative medicine GLP (Good Laboratory Practice), and includes three uniquely designed experiments. We provide cells after conducting equivalent tests in our own laboratory, and the results of verifying the quality of stem cells that are fairly provided to patients to third-party testing companies are also described here.
Adipose tissue from three sample donors was cultured at our laboratory and sent for inspection. The sample is taken from the back of the ear, and the tissue is separated and decomposed using special technology and a dedicated enzyme. Single cells are cultured for one month, and cells that reach a specified number are separated by MACS. After confirming the number of cells with a cell counter, they are provided to Takara Bio Inc., a third-party inspection company. bottom.
| Test purpose | exam name | Specification sample | result |
|---|---|---|---|
| infection, contamination | Sterility test (Japanese Pharmacopoeia Direct Method Option: Method Compatibility Test) |
QC_AL Lot:#13349 | negative |
| Sterility test (Japanese Pharmacopoeia direct method) |
QC_AL Lot:#13349 | negative | |
| Mycoplasma negative test (Japanese Pharmacopoeia reference information PCR method (confirmed alienation of 7 bacterial species)) |
QC_AL Lot:#13349 | negative | |
| endotoxin test (Japanese Pharmacopoeia Kinetic-Nephelometry Option: Reactive Dropping Factor Test) |
QC_AL Lot:#13349 | negative | |
| endotoxin test (Japanese Pharmacopoeia Kinetic - Nephelometry) |
QC_AL Lot:#13349 | Less than detection accuracy | |
| Tumorization | Colony formation test | QC_AL Lot:#13349 | negative |
| Stem cell confirmation | FCM exam (CD45-CD105+) |
QC_AL Lot:#13349 | CD45- 92.1% CD105+ 100% |
The above is an image diagram of adipose tissue, and the round object like a bean is the adipose tissue. Stem cells hide in a fibrous web-like area called the stem cell niche. In other words, stem cells are not present in lipid droplets, but are present in the surrounding fibrous places.
Let's take a look at the difference between ear fat and belly fat.
| ear fat | abdominal subcutaneous fat | |
| Appearance photo |  |
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| figure |  |
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| pain | few | strong (pain like a bruise) |
| internal bleeding | very rare | many |
| oppression | Band-aid pressure only | gauze compression |
Adipose-derived cells are not located in the yellow lipid droplets, but are lurking around the lipid droplets (= stem cell niche) called the stroma.
Stem cells are present in both abdominal fat and ear back fat, but the rate of stem cells is higher in "fat with less lipid droplets and more interstitium".
At Helene, we mainly collect behind-the-ear fat, which is less painful for the patient.
However, for those who prefer abdominal fat, abdominal liposuction can also be used.
Stem cells cultured at HELENE Cell Center are measured in at least two stages to confirm stem cell number and viability prior to dosing and delivery to customers. Results are lab approved and printed. We serve all our customers in a scientific and responsible way.
After the cell sample is obtained, staining is performed by adding 0.4% taipan blue solution, a vital stain commonly used to distinguish between live and dead cells. The staining method is a dye exclusion test, and the fact that the staining solution permeates dead cells but is blocked by the cell membrane of living cells is used to distinguish cells for viability calculation. Conventionally, stained samples were observed under a microscope and counted manually, but the automated cell automatically recognizes the fluoroscopic reaction and completes the analysis. For manual counting, not only must the focusing be done manually, but also the amount of brightfield light between samples must be adjusted. The Countess II FL Automated Cell Counter can automatically adjust lighting and focus for optimal image quality when in use.
After quantification with the Countess II FL, our cell center mounts the cell samples on slides and acquires cell images with the ZEISS Axio Vert.A1 microscope. HELENE's proprietary AI program for image reading was used to observe the cytoskeleton at the confocal level, combining the focal length of the microscope and the cell type (MSC) and the cross-sectional area of the cell culture flask to determine cell number, cell Calculate the survival rate. With these two steps, calculation accuracy is close to 100%, and an unrivaled artisanal process, HELENE brand therapeutic grade stem cells are precisely tailored to your needs and constantly optimized. We guarantee that
At HELENE Clinic, we demonstrate our commitment to quality and transparency by providing a Stem Cell Certificate for each treatment.
According to the 2021 paper [The safety of MSC therapy over the past 15 years: a meta-analysis], 62 studies including excerpts from clinical trials published in the internationally renowned journal "Lancet" were analyzed. is selected as Over 500 million, and in some cases as many as 1 billion, MSCs have been administered in these clinical trials, dispelling myths and confirming the technical capabilities of HELENE clinics alongside world-class research institutes.
| author | Year | Location | Dose | |
|---|---|---|---|---|
| Hess | 2017 | USA | 1200×106 cells | 1.2 billion stem cells |
| Bartunek | 2013 | Belgium | 600 -1200×106 cells | 6-1.2 billion stem cells |
| Chambers | 2017 | Australia | 1×109 cells | 1 billion stem cells / lung disorder |
| Steinberg | 2017 | USA | 1×109 cells | 1 billion stem cells / stroke |
| Igresias | 2021 | Mexico | 1×109 cells | 1 billion stem cells / lung disorder |
| Matsuoka | 2024 | Japan | 1-2×109 cells | 2 billion stem cells / AntiAging |
A large body of clinical data suggested that low doses of MSCs might not even be effective in some cases, as we were unable to find cases of low doses of cells for specific diseases. In the near future, with scientific data and research updates, we will continuously improve MSC treatment protocols, treatment targets, sources, culture protocols, dosage differences, etc. to better compare the results of relevant clinical trials. and should be standardized. Prior to this, HELENE Clinic has developed a re-infusion program with different amounts of cells and exosomes according to the patient's age, physical condition, and disease, and in order to guarantee the best stem cell treatment under complete safety, the treatment method continue to improve.
[Quote] The safety of MSC therapy over the past 15 years: a meta-analysis.
https://doi.org/10.1186/s13287-021-02609-x
| Consultation hours | 10:00～19:00 |
|---|---|
| Closed days | Wed/Sun |
| address | 〒107-0062 5-9-15 Minami Aoyama, Minato Ward, Tokyo Aoyama OHMOTO Building 3F |
| phone | 03-3400-2277 |