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CN-122012384-A - Preparation BMSCs-sec for repairing scar uterus and preparation method thereof

CN122012384ACN 122012384 ACN122012384 ACN 122012384ACN-122012384-A

Abstract

The invention discloses a preparation BMSCs-sec for repairing scar uterus and a preparation method thereof, which comprises the steps of carrying out passage on primary BMSCs with good growth to the third generation, collecting BMSC cells, inoculating the BMSCs into a culture flask, culturing in a complete culture medium, discarding the supernatant when the cell adhesion fusion rate reaches 80%, washing cells by PBS, adding DMEM/F12 for culturing, collecting the cell supernatant, centrifuging, removing cell fragments and dead cells, filtering by a filter, collecting filtrate, transferring the filtrate into a ultrafilter tube, centrifuging, and obtaining concentrated cell supernatant to obtain the preparation BMSCs-sec. The preparation BMSCs-sec prepared by the invention is used for repairing scar uterus, and improves a new thought for treating uterine scar injury by utilizing cell secretion.

Inventors

  • YANG JING
  • WEN YANQING
  • CHEN QIAN

Assignees

  • 贵州省人民医院
  • 重庆市妇幼保健院(重庆市妇产科医院、重庆市遗传与生殖研究所)

Dates

Publication Date
20260512
Application Date
20251231

Claims (9)

  1. 1. A preparation method of a preparation BMSCs-sec for repairing scar uterus is characterized by comprising the steps of, The primary bone marrow mesenchymal stem cells BMSCs with good growth are passaged to the third generation, and BMSC cells are collected; inoculating BMSCs into a culture flask, culturing in a complete culture medium, discarding the supernatant when the cell adhesion fusion rate reaches 80%, washing cells with PBS, adding DMEM/F12 for culturing, collecting cell supernatant, centrifuging, removing cell debris and dead cells, filtering by a filter, and collecting filtrate; Transferring the filtrate into a ultrafilter tube, centrifuging, and obtaining concentrated cell supernatant to obtain preparation BMSCs-sec.
  2. 2. The method of claim 1, wherein the primary BMSCs are derived from rat bone marrow.
  3. 3. The method of claim 1 or 2, wherein the DMEM/F12 is added for 24 to 48 hours.
  4. 4. The method of claim 3, wherein the centrifugation is performed at a rotational speed of 1000 Xg for 15 to 20 minutes.
  5. 5. The method according to claim 4, wherein the filter has a particle size of 0.22. Mu.m.
  6. 6. The method of claim 1 or 5, wherein the size of the ultrafiltration tube is 3K.
  7. 7. The method of claim 6, wherein the centrifugation is performed at a temperature of 4℃and a centrifugation speed of 5000 Xg to obtain a concentrated cell supernatant.
  8. 8. The BMSCs-sec preparation obtained by the production method according to any one of claims 1 to 7.
  9. 9. Use of the formulation BMSCs-sec of claim 8 for the manufacture of a medicament for repairing a scar uterus.

Description

Preparation BMSCs-sec for repairing scar uterus and preparation method thereof Technical Field The invention belongs to the technical field of biology, and particularly relates to a preparation BMSCs-sec for repairing scar uterus and a preparation method thereof. Background The scar uterus refers to uterus after caesarean section operation or myoma stripping operation, and is mainly caused by the defect of myometrium healing after operation, thereby causing uterine scar formation and related complications, and seriously affecting the life quality and fertility of patients. At present, the treatment of the uterine scar mainly comprises drug treatment, such as antiprogestin drug treatment, such as progesterone capsule and the like, for promoting the recovery of the uterine scar, and surgical treatment, such as hysteroscopic surgery and the like, for excising the uterine scar so as to achieve the purpose of recovering the normal uterine function. However, these traditional treatments have limited effectiveness, such as high recurrence rates for moderate and severe patients. Thus, there is a need for new and effective alternatives to repair and treat uterine scars, stem cell therapy being one of the major directions of development at present. However, most stem cells have weak proliferation and directional migration to endometrium, and cannot effectively play a role in repair. Currently, there is no report of BMSCs-derived secretome for repair treatment of uterine scars. Disclosure of Invention This section is intended to outline some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. The present invention has been made in view of the above and/or problems occurring in the prior art. Therefore, the invention aims to overcome the defects in the prior art and provide a preparation method of the preparation BMSCs-sec for repairing scar uterus. In order to solve the technical problems, the invention provides a preparation method of a preparation BMSCs-sec for repairing scar uterus, which comprises the following steps of, The well-grown primary BMSCs are passaged to the third generation, and BMSC cells are collected; inoculating BMSCs into a culture flask, culturing in a complete culture medium, discarding the supernatant when the cell adhesion fusion rate reaches 80%, washing cells with PBS, adding DMEM/F12 for culturing, collecting cell supernatant, centrifuging, removing cell debris and dead cells, filtering by a filter, and collecting filtrate; Transferring the filtrate into a ultrafilter tube, centrifuging, and obtaining concentrated cell supernatant to obtain preparation BMSCs-sec. As a preferred embodiment of the preparation method of the present invention, the primary BMSCs are derived from rat bone marrow. As a preferable scheme of the preparation method, the DMEM/F12 is added for 24-48 h for culture. As a preferable scheme of the preparation method, the method comprises the steps of centrifuging, removing cell fragments and dead cells, wherein the centrifugal speed is 1000 Xg, and the centrifugal time is 15-20 min. As a preferable mode of the production method of the present invention, wherein the filter is filtered, wherein the filter has a particle diameter of 0.22. Mu.m. As a preferable scheme of the preparation method, the specification of the ultrafiltration tube is 3K. As a preferred embodiment of the production method of the present invention, the centrifugation is performed at a temperature of 4℃and a rotational speed of 5000 Xg to obtain a concentrated cell supernatant. It is a further object of the present invention to overcome the deficiencies in the prior art and to provide a formulation of BMSCs-sec. The invention further aims to overcome the defects in the prior art and provide an application of BMSCs-sec in preparing medicines for repairing scar uterus. The invention has the beneficial effects that: The invention provides a preparation BMSCs-sec for repairing scar uterus, which promotes USMCs proliferation, migration and chemotaxis, reduces apoptosis and necrosis after USMCs injury, can regulate and control macrophage state, promote macrophage M2 polarization, promote angiogenesis, and can activate PI3K/AKT channel in USMCs, wherein secretory protein is a main bioactive component of BMSCs-sec regulation USMCs, and has good effect for repairing scar uterus; the preparation BMSCs-sec prepared by the invention is used for repairing scar uterus, and provides a new thought for treating uterine scar injury by utilizing cell secretion. Drawings In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the description of the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a