Protective solution and preparation method and application thereof

Abstract

The invention discloses a protective solution as well as a preparation method and application thereof. The protective solution comprises a cryoprotectant and a complete culture medium, the cryoprotectant is composed of a permeable protective agent and a non-permeable protective agent, and the non-permeable protective agent contains beta-1, 3 glucan; the complete culture medium consists of a serum-free basal culture medium, a serum substitute and L-glutamine; and the protective solution does not contain fetal bovine serum, autologous cord blood serum and human serum albumin. The protective solution can provide a good cryopreservation environment for tissue cells, beta-1, 3 glucan can improve the stability of the protective solution and improve the cryopreserved tissue protection effect andthe cell viability after resuscitation, the complete culture medium provides nutrition for umbilical cord tissues and cells and is consistent with a culture medium for stem cell resuscitation and culture, so that the convenience in preparation and application of the protective solution is improved. The protective solution has the advantages of being convenient to use, short in preparation period of a cryopreservation preparation, low in cost, safe and reliable in clinical application and the like.

Description

technical field [0001] The present application relates to the technical field of cryopreservation of tissue cells, in particular to a protective solution for cryopreservation of tissue cells and its preparation method and application. Background technique [0002] The umbilical cord is a tubular structure connecting the placenta and the fetus. The outer layer is amniotic membrane, and there are 2 umbilical arteries and 1 umbilical vein inside. Tong's jelly tissue. Through culture, about 10 6 primary mesenchymal stem cells. [0003] Umbilical cord mesenchymal stem cells are pluripotent stem cells with the ability of self-renewal and multi-directional differentiation, can differentiate into various cells and tissues, have low immunogenicity, can secrete various cytokines, regulate the immune system, and promote tissue and organ damage It can be used to treat osteoarthritis, graft-versus-host disease, diabetes and other diseases; therefore, umbilical cord Wharton's jelly tis...

AI Technical Summary

Problems solved by technology

For protective solutions containing fetal bovine serum or autologous cord blood serum, fetal bovine serum is of xenogeneic animal origin and may contain dangerous components such as undetected proteins or viruses, which will greatly increase the risk of clinical application of stem cells; while autologous cord blood The source of serum cannot be guaranteed, the amount collected is limited, and the quality of serum will be affected by various factors such as the current physical condition, so the quality cannot be guaranteed to be stable and reliable

In addition, there are batch differences between fetal bovine serum and autologous cord blood serum, which cannot guarantee the uniformity of quality

Therefore, the quality and stability of the protection solution cannot be guaranteed

For the protection solution containing human serum albumin, which is a human blood product and is an allogeneic component; when using cryopreservation solution containing human serum albumin to store tissues and cells, there will be certain transmission Potential Risk of Pathogens

Moreover, human serum albumin also has batch differences, which cannot guarantee the uniformity of quality

Moreover, the high cost of human albumin is not conducive to production cost control

[0006] In general, cryopreservation solutions in the prior art generally contain animal serum or human serum albumin, the preparation cycle of cryopreservation preparations is long, the preparation cost is high, and cell viability is damaged during cryopreservation and resuscitation, which is not conducive to the future clinical application of stem cells. There are many problems, and there is an urgent demand for safer, more reliable and low-cost cell and tissue cryopreservation technology in the industry

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