CN-121450582-B - Use of a composition, a culture medium additive or a culture medium for expanding hematopoietic stem cells or for preparing products for expanding hematopoietic stem cells

Abstract

The application provides the use of a composition, medium additive or medium for expanding hematopoietic stem cells or for preparing a product for expanding hematopoietic stem cells. In a first aspect the application provides a composition comprising quinizarine and resveratrol, a culture medium additive comprising a composition, a culture medium comprising a basal medium and an additive comprising a composition, or the use of a culture medium in the expansion of hematopoietic stem cells or in the preparation of a product for the expansion of hematopoietic stem cells. By screening specific HDAC inhibitors in combination with a variety of other small molecule compounds, the proportion of CD34 + 、CD34 + CD90 + and CD34 + CD90 + EPCR + cells after HSC expansion in vitro, i.e., hematopoietic stem cells and long-acting hematopoietic stem cells, can be significantly increased at lower working concentrations.

Inventors

• LI YUXI
• XU WENFEI
• HUA JUNNAN
• HU DINGFANG
• OUYANG WENJIE
• LIU CHAO
• GU YING

Assignees

• 深圳华大生命科学研究院
• 深圳市禾沐基因生物技术有限责任公司

Dates

Publication Date

20260505

Application Date

20260107

Claims (6)

1. 1. The application of the composition, the culture medium additive or the culture medium in the amplification of peripheral blood hematopoietic stem cells or the preparation of products for the amplification of peripheral blood hematopoietic stem cells for non-disease diagnosis or treatment purpose is characterized in that the composition comprises quininostat and resveratrol, the culture medium additive comprises the composition, the culture medium comprises a basal culture medium and an additive, the additive comprises the composition, the amplification product of the peripheral blood hematopoietic stem cells has long-acting hematopoietic stem cell CD34 + CD90 + EPCR + cells with higher proportion, wherein the working concentration of the quininostat is 0.5-5 nmol/L, and the working concentration of the resveratrol is 5-50 mu mol/L.
2. 2. The use according to claim 1, wherein the composition further comprises at least one of tranylcypromine and pomalidomide, wherein the working concentration of tranylcypromine is 1-10 μmol/L and the working concentration of pomalidomide is 1-10 μmol/L.
3. 3. The application of claim 1, wherein the composition comprises quinizarine, resveratrol and tranylcypromine, wherein the working concentration of quinizarine is 0.5-5 nmol/L, the working concentration of resveratrol is 5-50 mu mol/L, and the working concentration of tranylcypromine is 1-10 mu mol/L.
4. 4. The use according to claim 1, wherein the composition comprises quinizarine, resveratrol and pomalidomide, wherein the working concentration of quinizarine is 0.5-5 nmol/L, the working concentration of resveratrol is 5-50 μmol/L, and the working concentration of pomalidomide is 1-10 μmol/L.
5. 5. The use of claim 1, wherein the basal medium further comprises a cytokine, wherein the cytokine comprises at least one of a stem cytokine, an Fms-like tyrosine kinase 3 ligand, thrombopoietin, interleukin-6, interleukin-3.
6. 6. The use of claim 1, wherein said expanding peripheral blood hematopoietic stem cells comprises enriching CD34 + CD90 + EPCR + cells from an expanded product of said peripheral blood hematopoietic stem cells.

Description

Use of a composition, a culture medium additive or a culture medium for expanding hematopoietic stem cells or for preparing products for expanding hematopoietic stem cells Technical Field The application relates to the technical field of cell biology, in particular to application of a composition, a culture medium additive or a culture medium in expanding hematopoietic stem cells or preparing products for expanding the hematopoietic stem cells. Background Hematopoietic stem cells (Hematopoietic stem cell, HSCs) are stem cells located in hematopoietic tissues that have self-renewing and multipotent differentiation potential and are capable of differentiating into various types of blood cells, including erythrocytes, leukocytes, platelets, etc., thereby maintaining the normal functions of the blood system and immune system. Based on this principle, HSC transplantation is also an effective treatment for a variety of diseases including blood system diseases, immune system diseases, and some metabolic genetic diseases. Currently the major sources of HSCs include bone marrow, mobilized peripheral blood, and umbilical cord blood. Among them, the collection operation of the peripheral blood HSC is simple and patient-friendly, but the proportion of HSC in the peripheral blood is very low, and the injection of granulocyte colony stimulating factor (G-CSF) or praoxafu (Plerixafor) is required to mobilize hematopoietic stem cells in bone marrow into the peripheral blood, and there is a risk that repeated or even failure is required because of insufficient HSC. Clinical HSCs generally refer to CD34 + cells in the blood, which contain the most primitive hematopoietic stem cells (long-acting hematopoietic stem cells, LT-HSCs), short-term hematopoietic stem cells, multipotent progenitor cells, common myeloid progenitor cells, and common gonomic progenitor cells, among other cells with different self-renewal and differentiation potential, whereas only LT-HSCs can function for a long period of time and other stem cells are gradually depleted. LT-HSC are generally defined as cells of CD34 +CD38-CD45RA-CD90+, depending on the expression of the surface protein. However, it was found that CD38 + cells lost CD38 expression during in vitro culture and thus CD38 was not suitable for characterizing in vitro cultured HSCs. The proportion of LT-HSC in the CD34 +CD90+ cell population is higher than that in the CD34 + cells, and the proportion is still lower. In recent years, EPCR has been found to be a good characterization of in vitro expanded LT-HSCs, which are highly enriched in the CD34 +CD90+EPCR+ group. HSC transplantation effects are closely related to the number of transplanted cells, and when the number of transplanted HSCs is small, it takes longer to reconstitute the blood and immune system, and the risk of infection increases. In addition to collecting more cells, in vitro expansion of HSCs is also an effective means to address clinical cell sources and numbers shortages and improve hematopoietic stem cell engraftment success rates. Related studies have attempted a number of different approaches to achieve in vitro expansion of HSCs, with the fastest clinical progression being the addition of specific small molecule compounds to in vitro cultures of HSCs, thereby facilitating expansion and maintenance of HSCs. For example, mobilized peripheral blood purified CD34 + cells were cultured in an in vitro culture procedure with 1mM VPA, but the results found that the proportion of CD34 + cells in the expanded cells was low and the proportion of expanded long-acting hematopoietic stem cells could not be accurately characterized. And its higher addition concentration may introduce more chemical residues in the cell product, with a certain safety risk. Accordingly, there is a need to provide a composition that can be expanded to yield HSCs with higher LT-HSC ratios at lower working concentrations. Disclosure of Invention The main purpose of the embodiment of the invention is to provide an application of a composition, a culture medium additive or a culture medium in the expansion of hematopoietic stem cells or the preparation of products for the expansion of hematopoietic stem cells, which can be used for expanding and obtaining HSC with higher LT-HSC ratio under lower working concentration. To achieve the above object, the first aspect of the present application provides a composition, a culture medium additive or a use of a culture medium in expanding hematopoietic stem cells or preparing products for expanding hematopoietic stem cells, the composition comprising quininostat and resveratrol, the culture medium additive comprising a composition, the culture medium comprising a basal medium and an additive, the additive comprising a composition. Wherein, quininostat (Quisinostat) is an inhibitor of ubiquitin deacetylase (HDAC), CAS number 875320-29-9, and has the following structural formula: 。 Resveratrol is a non-flavonoid polyphe