**Part 2**

**Human Asisited Reproduction Techniques (ART)** 

102 Current Frontiers in Cryopreservation

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Marrale, J; Morrissey, MC; Haddad, FS. (2007)A literature review of autograft and allograft

Nutton, RW; McLean, I; Melville, E. (1999) Tendon allografts in knee ligament surgery. *J R* 

Papandrea, P; Vulpiani , MC; Ferretti, A; Conteduca, F. (2000) Regeneration of the

Rappe, M; Horodyski, M; Meister, K; Indelicato, PA. (2007) Nonirradiated versus irradiated

Reiff, RBM; Croci, AT; Bolliger Neto, R; Pereira, CAM. (2007) Estudo comparativo de

frozen and fresh calcaneus tendon]. *Acta Ortop Bras,* 15, 6-8. ISSN 2176-7521. Schachar, NS; McGann, LE. (1986) Investigations of low-temperature storage of articular cartilage for transplantation. *Clin Orthop Relat Res,* 146- 50. ISSN 0009-921X. Sherman, OH; Banffy, MB. (2004) Anterior cruciate ligament reconstruction: which graft is

Sterling, JC; Meyers, MC; Calvo RD. (1995) Allograft failure in cruciate ligament

Sun, K; Zhang, J; Wang, Y; Xia, C; Zhang, C; Yu, T; Tian, S. (2011) Arthroscopic

Tomford, W. Transmission of disease through musculoskeletal transplantation. *Portland Bone Symposium.* Portland: Oregon Health Sciences University, 1997. Urabe, K; Itoman, M; Toyama, Y; Yanase, Y; Iwamoto, Y; Ohgushi, H. (2007) Current trends

Vangsness, CT Jr; Garcia, IA; Mills, CR; Kainer, MA; Roberts, MR; Moore, TM.

processing, and sterilization. *Am J Sports Med,* 31,474-481. ISSN 1552-3365. Zimmerman, MC; Contiliano, JH; Parsons, JR; Prewett, A; Billotti, J. (1994) The biomechanics

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*Arthroscopy*, 19, 1091-6. ISSN 0749-8063.

best? *Arthroscopy,* 20, 974-80. ISSN 0749-8063.

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*Coll Surg Edinb,* 44, 236-40.

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8. ISSN 1552-3365.

525. ISSN 1436-2023.

ligaments with bone-patellar tendon-bone and achilles tendon allografts. *Clin* 

anterior cruciate ligament reconstruction. *Knee Surg Sports Traumatol Arthrosc,* 15,

semitendinosus tendon harvested for anterior cruciate ligament reconstruction. Evaluation using ultrasonography. *Am J Sports Med*, 28, 556-61. ISSN 1552-3365. Pearsall, AW; Hollis, JM; Russell, GV Jr; Scheer, Z. (2003) A biomechanical comparison of

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**6**

*Spain* 

**The Problem of Contamination:**

*CERAM: Centre for assisted reproduction in Marbella* 

 **Vitrification Systems** 

Enrique Criado Scholz

**Open** *vs.* **Closed** *vs.* **Semi-Closed** 

The Development of cryopreservation techniques, the increase in demand for cryopreserved cells or tissues and the use of these techniques in cells or tissues from patients with infectious diseases, has forced us to reduce the risk of contamination during the freezing process and the risk of cross-contamination during the storage of this material. Recent publications that demonstrate the survival of pathogens at low temperatures and possible contamination of the cells or tissues stored have changed the laws of each country and the

To understand the problem of contamination in cryopreservation we need to have an overview of the current problem in which all researchers are concerned about, seeking a cryopreservation protocol with good results but without contamination problems. Discussing the cryopreservation's different techniques such as slow freezing, vitrification, kinetic vitrification (extra-, hyper-, super-, ultra-fast vitrification) and the various components that help us understand the difficult balance between technique, device used and the risk of contamination. We need to use new products and new protocols' to have

Dr. Katkov's idea of find the *"Universal crypreservation protocol"* (see the other Chapter in the Book) by Katkov at al. that we can use worldwide with all the possible biological samples would lower considerably the price of cryopreservation process and we would have better

We have to comprehend the difference between open, closed or semi-closed devices and the importance of choosing one device or another both in morphological survival post thaw cell or tissue as on non-contamination of these samples. The device used, the protocol used and the cooling solution used can change the outcome of cryopreservation and therefore we have to find a protocol for cryopreservation with a cooling solution and a secure device to

The first thing we must learn is to differentiate their respective importance are the concepts of contamination and cross contamination of samples. The first relates to the contamination

results because everybody would work with the same protocol and the same results.

customs and protocols used so far in the cryopreservation.

good results ensuring biological samples and patient safety.

provide us good results free of contamination.

**2. Contamination and cross-contamination** 

**1. Introduction** 
